Publisher: Elsevier   (Total: 3147 journals)

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Showing 1 - 200 of 3147 Journals sorted alphabetically
Academic Pediatrics     Hybrid Journal   (Followers: 39, SJR: 1.655, CiteScore: 2)
Academic Radiology     Hybrid Journal   (Followers: 26, SJR: 1.015, CiteScore: 2)
Accident Analysis & Prevention     Partially Free   (Followers: 106, SJR: 1.462, CiteScore: 3)
Accounting Forum     Hybrid Journal   (Followers: 28, SJR: 0.932, CiteScore: 2)
Accounting, Organizations and Society     Hybrid Journal   (Followers: 44, SJR: 1.771, CiteScore: 3)
Achievements in the Life Sciences     Open Access   (Followers: 7)
Acta Anaesthesiologica Taiwanica     Open Access   (Followers: 6)
Acta Astronautica     Hybrid Journal   (Followers: 447, SJR: 0.758, CiteScore: 2)
Acta Automatica Sinica     Full-text available via subscription   (Followers: 2)
Acta Biomaterialia     Hybrid Journal   (Followers: 30, SJR: 1.967, CiteScore: 7)
Acta Colombiana de Cuidado Intensivo     Full-text available via subscription   (Followers: 3)
Acta de Investigación Psicológica     Open Access   (Followers: 2)
Acta Ecologica Sinica     Open Access   (Followers: 12, SJR: 0.18, CiteScore: 1)
Acta Histochemica     Hybrid Journal   (Followers: 5, SJR: 0.661, CiteScore: 2)
Acta Materialia     Hybrid Journal   (Followers: 325, SJR: 3.263, CiteScore: 6)
Acta Mathematica Scientia     Full-text available via subscription   (Followers: 5, SJR: 0.504, CiteScore: 1)
Acta Mechanica Solida Sinica     Full-text available via subscription   (Followers: 9, SJR: 0.542, CiteScore: 1)
Acta Oecologica     Hybrid Journal   (Followers: 12, SJR: 0.834, CiteScore: 2)
Acta Otorrinolaringologica (English Edition)     Full-text available via subscription  
Acta Otorrinolaringológica Española     Full-text available via subscription   (Followers: 2, SJR: 0.307, CiteScore: 0)
Acta Pharmaceutica Sinica B     Open Access   (Followers: 2, SJR: 1.793, CiteScore: 6)
Acta Psychologica     Hybrid Journal   (Followers: 26, SJR: 1.331, CiteScore: 2)
Acta Sociológica     Open Access   (Followers: 1)
Acta Tropica     Hybrid Journal   (Followers: 6, SJR: 1.052, CiteScore: 2)
Acta Urológica Portuguesa     Open Access   (Followers: 1)
Actas Dermo-Sifiliograficas     Full-text available via subscription   (Followers: 3, SJR: 0.374, CiteScore: 1)
Actas Dermo-Sifiliográficas (English Edition)     Full-text available via subscription   (Followers: 2)
Actas Urológicas Españolas     Full-text available via subscription   (Followers: 3, SJR: 0.344, CiteScore: 1)
Actas Urológicas Españolas (English Edition)     Full-text available via subscription   (Followers: 1)
Actualites Pharmaceutiques     Full-text available via subscription   (Followers: 7, SJR: 0.19, CiteScore: 0)
Actualites Pharmaceutiques Hospitalieres     Full-text available via subscription   (Followers: 3)
Acupuncture and Related Therapies     Hybrid Journal   (Followers: 8)
Acute Pain     Full-text available via subscription   (Followers: 15, SJR: 2.671, CiteScore: 5)
Ad Hoc Networks     Hybrid Journal   (Followers: 11, SJR: 0.53, CiteScore: 4)
Addictive Behaviors     Hybrid Journal   (Followers: 18, SJR: 1.29, CiteScore: 3)
Addictive Behaviors Reports     Open Access   (Followers: 9, SJR: 0.755, CiteScore: 2)
Additive Manufacturing     Hybrid Journal   (Followers: 13, SJR: 2.611, CiteScore: 8)
Additives for Polymers     Full-text available via subscription   (Followers: 22)
Advanced Drug Delivery Reviews     Hybrid Journal   (Followers: 190, SJR: 4.09, CiteScore: 13)
Advanced Engineering Informatics     Hybrid Journal   (Followers: 13, SJR: 1.167, CiteScore: 4)
Advanced Powder Technology     Hybrid Journal   (Followers: 17, SJR: 0.694, CiteScore: 3)
Advances in Accounting     Hybrid Journal   (Followers: 9, SJR: 0.277, CiteScore: 1)
Advances in Agronomy     Full-text available via subscription   (Followers: 17, SJR: 2.384, CiteScore: 5)
Advances in Anesthesia     Full-text available via subscription   (Followers: 30, SJR: 0.126, CiteScore: 0)
Advances in Antiviral Drug Design     Full-text available via subscription   (Followers: 2)
Advances in Applied Mathematics     Full-text available via subscription   (Followers: 12, SJR: 0.992, CiteScore: 1)
Advances in Applied Mechanics     Full-text available via subscription   (Followers: 12, SJR: 1.551, CiteScore: 4)
Advances in Applied Microbiology     Full-text available via subscription   (Followers: 24, SJR: 2.089, CiteScore: 5)
Advances In Atomic, Molecular, and Optical Physics     Full-text available via subscription   (Followers: 15, SJR: 0.572, CiteScore: 2)
Advances in Biological Regulation     Hybrid Journal   (Followers: 4, SJR: 2.61, CiteScore: 7)
Advances in Botanical Research     Full-text available via subscription   (Followers: 1, SJR: 0.686, CiteScore: 2)
Advances in Cancer Research     Full-text available via subscription   (Followers: 35, SJR: 3.043, CiteScore: 6)
Advances in Carbohydrate Chemistry and Biochemistry     Full-text available via subscription   (Followers: 9, SJR: 1.453, CiteScore: 2)
Advances in Catalysis     Full-text available via subscription   (Followers: 5, SJR: 1.992, CiteScore: 5)
Advances in Cell Aging and Gerontology     Full-text available via subscription   (Followers: 5)
Advances in Cellular and Molecular Biology of Membranes and Organelles     Full-text available via subscription   (Followers: 14)
Advances in Chemical Engineering     Full-text available via subscription   (Followers: 29, SJR: 0.156, CiteScore: 1)
Advances in Child Development and Behavior     Full-text available via subscription   (Followers: 11, SJR: 0.713, CiteScore: 1)
Advances in Chronic Kidney Disease     Full-text available via subscription   (Followers: 11, SJR: 1.316, CiteScore: 2)
Advances in Clinical Chemistry     Full-text available via subscription   (Followers: 26, SJR: 1.562, CiteScore: 3)
Advances in Colloid and Interface Science     Full-text available via subscription   (Followers: 21, SJR: 1.977, CiteScore: 8)
Advances in Computers     Full-text available via subscription   (Followers: 14, SJR: 0.205, CiteScore: 1)
Advances in Dermatology     Full-text available via subscription   (Followers: 16)
Advances in Developmental Biology     Full-text available via subscription   (Followers: 14)
Advances in Digestive Medicine     Open Access   (Followers: 13)
Advances in DNA Sequence-Specific Agents     Full-text available via subscription   (Followers: 7)
Advances in Drug Research     Full-text available via subscription   (Followers: 26)
Advances in Ecological Research     Full-text available via subscription   (Followers: 45, SJR: 2.524, CiteScore: 4)
Advances in Engineering Software     Hybrid Journal   (Followers: 30, SJR: 1.159, CiteScore: 4)
Advances in Experimental Biology     Full-text available via subscription   (Followers: 9)
Advances in Experimental Social Psychology     Full-text available via subscription   (Followers: 51, SJR: 5.39, CiteScore: 8)
Advances in Exploration Geophysics     Full-text available via subscription   (Followers: 2)
Advances in Fluorine Science     Full-text available via subscription   (Followers: 9)
Advances in Food and Nutrition Research     Full-text available via subscription   (Followers: 68, SJR: 0.591, CiteScore: 2)
Advances in Fuel Cells     Full-text available via subscription   (Followers: 17)
Advances in Genetics     Full-text available via subscription   (Followers: 21, SJR: 1.354, CiteScore: 4)
Advances in Genome Biology     Full-text available via subscription   (Followers: 12, SJR: 12.74, CiteScore: 13)
Advances in Geophysics     Full-text available via subscription   (Followers: 8, SJR: 1.193, CiteScore: 3)
Advances in Heat Transfer     Full-text available via subscription   (Followers: 26, SJR: 0.368, CiteScore: 1)
Advances in Heterocyclic Chemistry     Full-text available via subscription   (Followers: 11, SJR: 0.749, CiteScore: 3)
Advances in Human Factors/Ergonomics     Full-text available via subscription   (Followers: 26)
Advances in Imaging and Electron Physics     Full-text available via subscription   (Followers: 4, SJR: 0.193, CiteScore: 0)
Advances in Immunology     Full-text available via subscription   (Followers: 37, SJR: 4.433, CiteScore: 6)
Advances in Inorganic Chemistry     Full-text available via subscription   (Followers: 10, SJR: 1.163, CiteScore: 2)
Advances in Insect Physiology     Full-text available via subscription   (Followers: 2, SJR: 1.938, CiteScore: 3)
Advances in Integrative Medicine     Hybrid Journal   (Followers: 6, SJR: 0.176, CiteScore: 0)
Advances in Intl. Accounting     Full-text available via subscription   (Followers: 3)
Advances in Life Course Research     Hybrid Journal   (Followers: 9, SJR: 0.682, CiteScore: 2)
Advances in Lipobiology     Full-text available via subscription   (Followers: 1)
Advances in Magnetic and Optical Resonance     Full-text available via subscription   (Followers: 8)
Advances in Marine Biology     Full-text available via subscription   (Followers: 21, SJR: 0.88, CiteScore: 2)
Advances in Mathematics     Full-text available via subscription   (Followers: 17, SJR: 3.027, CiteScore: 2)
Advances in Medical Sciences     Hybrid Journal   (Followers: 9, SJR: 0.694, CiteScore: 2)
Advances in Medicinal Chemistry     Full-text available via subscription   (Followers: 6)
Advances in Microbial Physiology     Full-text available via subscription   (Followers: 5, SJR: 1.158, CiteScore: 3)
Advances in Molecular and Cell Biology     Full-text available via subscription   (Followers: 26)
Advances in Molecular and Cellular Endocrinology     Full-text available via subscription   (Followers: 8)
Advances in Molecular Toxicology     Full-text available via subscription   (Followers: 7, SJR: 0.182, CiteScore: 0)
Advances in Nanoporous Materials     Full-text available via subscription   (Followers: 5)
Advances in Oncobiology     Full-text available via subscription   (Followers: 2)
Advances in Organ Biology     Full-text available via subscription   (Followers: 2)
Advances in Organometallic Chemistry     Full-text available via subscription   (Followers: 18, SJR: 1.875, CiteScore: 4)
Advances in Parallel Computing     Full-text available via subscription   (Followers: 7, SJR: 0.174, CiteScore: 0)
Advances in Parasitology     Full-text available via subscription   (Followers: 6, SJR: 1.579, CiteScore: 4)
Advances in Pediatrics     Full-text available via subscription   (Followers: 27, SJR: 0.461, CiteScore: 1)
Advances in Pharmaceutical Sciences     Full-text available via subscription   (Followers: 19)
Advances in Pharmacology     Full-text available via subscription   (Followers: 17, SJR: 1.536, CiteScore: 3)
Advances in Physical Organic Chemistry     Full-text available via subscription   (Followers: 10, SJR: 0.574, CiteScore: 1)
Advances in Phytomedicine     Full-text available via subscription  
Advances in Planar Lipid Bilayers and Liposomes     Full-text available via subscription   (Followers: 3, SJR: 0.109, CiteScore: 1)
Advances in Plant Biochemistry and Molecular Biology     Full-text available via subscription   (Followers: 11)
Advances in Plant Pathology     Full-text available via subscription   (Followers: 6)
Advances in Porous Media     Full-text available via subscription   (Followers: 5)
Advances in Protein Chemistry     Full-text available via subscription   (Followers: 19)
Advances in Protein Chemistry and Structural Biology     Full-text available via subscription   (Followers: 20, SJR: 0.791, CiteScore: 2)
Advances in Psychology     Full-text available via subscription   (Followers: 69)
Advances in Quantum Chemistry     Full-text available via subscription   (Followers: 7, SJR: 0.371, CiteScore: 1)
Advances in Radiation Oncology     Open Access   (Followers: 3, SJR: 0.263, CiteScore: 1)
Advances in Small Animal Medicine and Surgery     Hybrid Journal   (Followers: 3, SJR: 0.101, CiteScore: 0)
Advances in Space Biology and Medicine     Full-text available via subscription   (Followers: 7)
Advances in Space Research     Full-text available via subscription   (Followers: 432, SJR: 0.569, CiteScore: 2)
Advances in Structural Biology     Full-text available via subscription   (Followers: 6)
Advances in Surgery     Full-text available via subscription   (Followers: 13, SJR: 0.555, CiteScore: 2)
Advances in the Study of Behavior     Full-text available via subscription   (Followers: 37, SJR: 2.208, CiteScore: 4)
Advances in Veterinary Medicine     Full-text available via subscription   (Followers: 20)
Advances in Veterinary Science and Comparative Medicine     Full-text available via subscription   (Followers: 15)
Advances in Virus Research     Full-text available via subscription   (Followers: 6, SJR: 2.262, CiteScore: 5)
Advances in Water Resources     Hybrid Journal   (Followers: 57, SJR: 1.551, CiteScore: 3)
Aeolian Research     Hybrid Journal   (Followers: 6, SJR: 1.117, CiteScore: 3)
Aerospace Science and Technology     Hybrid Journal   (Followers: 396, SJR: 0.796, CiteScore: 3)
AEU - Intl. J. of Electronics and Communications     Hybrid Journal   (Followers: 8, SJR: 0.42, CiteScore: 2)
African J. of Emergency Medicine     Open Access   (Followers: 6, SJR: 0.296, CiteScore: 0)
Ageing Research Reviews     Hybrid Journal   (Followers: 12, SJR: 3.671, CiteScore: 9)
Aggression and Violent Behavior     Hybrid Journal   (Followers: 490, SJR: 1.238, CiteScore: 3)
Agri Gene     Hybrid Journal   (Followers: 1, SJR: 0.13, CiteScore: 0)
Agricultural and Forest Meteorology     Hybrid Journal   (Followers: 18, SJR: 1.818, CiteScore: 5)
Agricultural Systems     Hybrid Journal   (Followers: 32, SJR: 1.156, CiteScore: 4)
Agricultural Water Management     Hybrid Journal   (Followers: 47, SJR: 1.272, CiteScore: 3)
Agriculture and Agricultural Science Procedia     Open Access   (Followers: 4)
Agriculture and Natural Resources     Open Access   (Followers: 3)
Agriculture, Ecosystems & Environment     Hybrid Journal   (Followers: 58, SJR: 1.747, CiteScore: 4)
Ain Shams Engineering J.     Open Access   (Followers: 5, SJR: 0.589, CiteScore: 3)
Air Medical J.     Hybrid Journal   (Followers: 8, SJR: 0.26, CiteScore: 0)
AKCE Intl. J. of Graphs and Combinatorics     Open Access   (SJR: 0.19, CiteScore: 0)
Alcohol     Hybrid Journal   (Followers: 12, SJR: 1.153, CiteScore: 3)
Alcoholism and Drug Addiction     Open Access   (Followers: 12)
Alergologia Polska : Polish J. of Allergology     Full-text available via subscription   (Followers: 1)
Alexandria Engineering J.     Open Access   (Followers: 2, SJR: 0.604, CiteScore: 3)
Alexandria J. of Medicine     Open Access   (Followers: 1, SJR: 0.191, CiteScore: 1)
Algal Research     Partially Free   (Followers: 11, SJR: 1.142, CiteScore: 4)
Alkaloids: Chemical and Biological Perspectives     Full-text available via subscription   (Followers: 2)
Allergologia et Immunopathologia     Full-text available via subscription   (Followers: 1, SJR: 0.504, CiteScore: 1)
Allergology Intl.     Open Access   (Followers: 5, SJR: 1.148, CiteScore: 2)
Alpha Omegan     Full-text available via subscription   (SJR: 3.521, CiteScore: 6)
ALTER - European J. of Disability Research / Revue Européenne de Recherche sur le Handicap     Full-text available via subscription   (Followers: 11, SJR: 0.201, CiteScore: 1)
Alzheimer's & Dementia     Hybrid Journal   (Followers: 55, SJR: 4.66, CiteScore: 10)
Alzheimer's & Dementia: Diagnosis, Assessment & Disease Monitoring     Open Access   (Followers: 6, SJR: 1.796, CiteScore: 4)
Alzheimer's & Dementia: Translational Research & Clinical Interventions     Open Access   (Followers: 6, SJR: 1.108, CiteScore: 3)
Ambulatory Pediatrics     Hybrid Journal   (Followers: 5)
American Heart J.     Hybrid Journal   (Followers: 58, SJR: 3.267, CiteScore: 4)
American J. of Cardiology     Hybrid Journal   (Followers: 67, SJR: 1.93, CiteScore: 3)
American J. of Emergency Medicine     Hybrid Journal   (Followers: 48, SJR: 0.604, CiteScore: 1)
American J. of Geriatric Pharmacotherapy     Full-text available via subscription   (Followers: 13)
American J. of Geriatric Psychiatry     Hybrid Journal   (Followers: 15, SJR: 1.524, CiteScore: 3)
American J. of Human Genetics     Hybrid Journal   (Followers: 40, SJR: 7.45, CiteScore: 8)
American J. of Infection Control     Hybrid Journal   (Followers: 29, SJR: 1.062, CiteScore: 2)
American J. of Kidney Diseases     Hybrid Journal   (Followers: 37, SJR: 2.973, CiteScore: 4)
American J. of Medicine     Hybrid Journal   (Followers: 50)
American J. of Medicine Supplements     Full-text available via subscription   (Followers: 3, SJR: 1.967, CiteScore: 2)
American J. of Obstetrics and Gynecology     Hybrid Journal   (Followers: 266, SJR: 2.7, CiteScore: 4)
American J. of Ophthalmology     Hybrid Journal   (Followers: 67, SJR: 3.184, CiteScore: 4)
American J. of Ophthalmology Case Reports     Open Access   (Followers: 5, SJR: 0.265, CiteScore: 0)
American J. of Orthodontics and Dentofacial Orthopedics     Full-text available via subscription   (Followers: 6, SJR: 1.289, CiteScore: 1)
American J. of Otolaryngology     Hybrid Journal   (Followers: 25, SJR: 0.59, CiteScore: 1)
American J. of Pathology     Hybrid Journal   (Followers: 32, SJR: 2.139, CiteScore: 4)
American J. of Preventive Medicine     Hybrid Journal   (Followers: 30, SJR: 2.164, CiteScore: 4)
American J. of Surgery     Hybrid Journal   (Followers: 39, SJR: 1.141, CiteScore: 2)
American J. of the Medical Sciences     Hybrid Journal   (Followers: 12, SJR: 0.767, CiteScore: 1)
Ampersand : An Intl. J. of General and Applied Linguistics     Open Access   (Followers: 7)
Anaerobe     Hybrid Journal   (Followers: 4, SJR: 1.144, CiteScore: 3)
Anaesthesia & Intensive Care Medicine     Full-text available via subscription   (Followers: 67, SJR: 0.138, CiteScore: 0)
Anaesthesia Critical Care & Pain Medicine     Full-text available via subscription   (Followers: 25, SJR: 0.411, CiteScore: 1)
Anales de Cirugia Vascular     Full-text available via subscription   (Followers: 1)
Anales de Pediatría     Full-text available via subscription   (Followers: 3, SJR: 0.277, CiteScore: 0)
Anales de Pediatría (English Edition)     Full-text available via subscription  
Anales de Pediatría Continuada     Full-text available via subscription  
Analytic Methods in Accident Research     Hybrid Journal   (Followers: 6, SJR: 4.849, CiteScore: 10)
Analytica Chimica Acta     Hybrid Journal   (Followers: 44, SJR: 1.512, CiteScore: 5)
Analytica Chimica Acta : X     Open Access  
Analytical Biochemistry     Hybrid Journal   (Followers: 216, SJR: 0.633, CiteScore: 2)
Analytical Chemistry Research     Open Access   (Followers: 13, SJR: 0.411, CiteScore: 2)
Analytical Spectroscopy Library     Full-text available via subscription   (Followers: 14)
Anesthésie & Réanimation     Full-text available via subscription   (Followers: 2)
Anesthesiology Clinics     Full-text available via subscription   (Followers: 25, SJR: 0.683, CiteScore: 2)
Angiología     Full-text available via subscription   (SJR: 0.121, CiteScore: 0)
Angiologia e Cirurgia Vascular     Open Access   (Followers: 1, SJR: 0.111, CiteScore: 0)
Animal Behaviour     Hybrid Journal   (Followers: 239, SJR: 1.58, CiteScore: 3)
Animal Feed Science and Technology     Hybrid Journal   (Followers: 8, SJR: 0.937, CiteScore: 2)
Animal Reproduction Science     Hybrid Journal   (Followers: 7, SJR: 0.704, CiteScore: 2)
Annales d'Endocrinologie     Full-text available via subscription   (Followers: 3, SJR: 0.451, CiteScore: 1)

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Similar Journals
Journal Cover
Analytical Biochemistry
Journal Prestige (SJR): 0.633
Citation Impact (citeScore): 2
Number of Followers: 216  
 
  Hybrid Journal Hybrid journal (It can contain Open Access articles)
ISSN (Print) 0003-2697 - ISSN (Online) 1096-0309
Published by Elsevier Homepage  [3147 journals]
  • Multi-analyses of gallstones and correlation between their properties with
           the laboratory results
    • Abstract: Publication date: 15 March 2020Source: Analytical Biochemistry, Volume 593Author(s): Anca Peter, Leonard Mihaly Cozmuta, Camelia Nicula, Anca Mihaly Cozmuța, Adriana Vulpoi, Lucian Barbu-Tudoran, Klara Magyari, Milica Todea, Lucian Baia, Flaviu Gheorghe PopThis study explores the morpho-structure of gallstones (GSs) removed from 36 patients in NW Romania and correlate it with the laboratory results of the patients. GSs were analyzed by SEM-EDS, X-ray diffraction and IR, UV–Vis and X-ray photoelectron spectroscopy. The laboratory studies consisted in hematological, coagulation, biochemistry, immunological and tumor markers tests. The morphological and structural investigations allowed to classify the GS in five different types and to establish their mechanism of formation. Only macroscopic evaluation, SEM microscopy, FTIR and UV–Vis spectroscopy give different easily noticeable information for each GS type. EDS, XPS and XRD diffraction are recommended to distinguish pigment and carbonate stones from the other GS types and a carefully examination is needed to establish the differences between the pure cholesterol, the mixed cholesterol and the composite cholesterol stones, due to the high similarities. The variation of specific markers cannot differentiate the patients with pure cholesterol GS from those with mixed cholesterol and pigment GS and those with mixed cholesterol from those with composite cholesterol stones. Seven laboratory parameters (RDW-CV, MPV, PCT, GLUC-HK, WBC, PT, GPT) are the key indicators for the GS disease and trend to present generally higher values than normal.Graphical abstractImage 1
       
  • Nicotinamide phosphoribosyltransferase purification using SUMO expression
           system
    • Abstract: Publication date: Available online 23 January 2020Source: Analytical BiochemistryAuthor(s): Trivikram R. Molugu, Radu C. Oita, Udeep Chawla, Sara M. Camp, Michael F. Brown, Joe G.N. GarciaNicotinamide phosphoribosyltransferase (NAMPT) is a rate-limiting enzyme in the salvage pathway required for nicotinamide adenine dinucleotide synthesis. The secreted NAMPT protein serves as a master regulatory cytokine involved in activation of evolutionarily-conserved inflammatory networks. Appreciation of the role of NAMPT as a damage-associated molecular pattern protein (DAMP) has linked its activities to several disorders via toll-like receptor 4 (TLR4) binding and inflammatory cascade activation. Information is currently lacking concerning the precise mode of the NAMPT protein functionality due to limited availability of purified protein for use in in vitro and in vivo studies. Here we report successful NAMPT expression using the pET-SUMO expression vector in E. coli strain SHuffle containing a hexa-His tag for purification. The Ulp1 protease was used to cleave the SUMO and hexa-His tags, and the protein was purified by immobilized-metal affinity chromatography. The protein yield was ∼4 mg/L and initial biophysical characterization of the protein using circular dichroism revealed the secondary structural elements, while dynamic light scattering demonstrated the presence of oligomeric units. The NAMPT-SUMO showed a predominantly dimeric protein with functional enzymatic activity. Finally, we report NAMPT solubilization in n-dodecyl-β-d-maltopyranoside (DDM) detergent in monomeric form, thus enhancing the opportunity for further structural and functional investigations.Graphical abstractImage 1
       
  • Identification of the epitope of 10-7G glycan antibody to recognize
           cancer-associated haptoglobin
    • Abstract: Publication date: Available online 22 January 2020Source: Analytical BiochemistryAuthor(s): Koichi Morishita, Yuta Maki, Shinji Takamatsu, Nami Ito, Sayaka Koda, Kei Motooka, Yoshihiro Kamada, Yasuhiro Kajihara, Eiji MiyoshiWe previously identified fucosylated haptoglobin (Fuc-Hpt) as a clinical serum biomarker of pancreatic cancer and established the novel glycan monoclonal antibody (mAb) 10-7G. This antibody recognizes cancer-associated haptoglobin including Fuc-Hpt and the precursor of haptoglobin. Interestingly, Western blot analysis showed that the 10-7G mAb reacts with the haptoglobin α chain, which has no N-glycan potential sites; haptoglobin β chain has 4 N-glycan sites. In this study, we identified the epitope for the 10-7G mAb using haptoglobin deletion mutants, as well as inhibition ELISA with recombinant peptides. We illustrated molecular graphics to show a relationship between the epitope and the β chain. Furthermore, we hypothesized that the 10-7G mAb minimally recognizes normal haptoglobin, but aberrant glycosylation on the β chain causes conformational changes, enabling the 10-7G mAb to easily access the epitope within the α chain. Because 10-7G values, an enzyme-linked immunosorbent assay–immobilized 10-7G mAb, in patients with pancreatic cancer varied by haptoglobin phenotype, the amount of aberrant glycosylation needed to affect haptoglobin conformation probably depends on haptoglobin phenotype. Taken together, the 10-7G mAb recognized characteristic peptides on the haptoglobin α chain as a result of conformational changes and is a promising tool for diagnosing pancreatic cancer by haptoglobin phenotype.Graphical abstractImage 1
       
  • Nanograss sensor for selective detection of Pseudomonas aeruginosa by
           pyocyanin identification in airway samples
    • Abstract: Publication date: Available online 22 January 2020Source: Analytical BiochemistryAuthor(s): Fatima AlZahra'a Alatraktchi, Maria Dimaki, Nicolai Støvring, Helle Krogh Johansen, Søren Molin, Winnie E. SvendsenPyocyanin is a virulence factor solely produced by the pathogen Pseudomonas aeruginosa. Pyocyanin is also a redox active molecule that can be directly detected by electrochemical sensing. A nanograss (NG) based sensor for sensitive quantification of pyocyanin in sputum samples from cystic fibrosis (CF) patients is presented here. The NG sensors were custom made in a cleanroom environment by etching nanograss topography on the electrode surface followed by depositing 200 nm gold. The NG sensors were utilized for amperometric quantification of pyocyanin in spiked hypertonic saline samples, resulting in a linear calibration curve with a R2 value of 0.9901 and a limit of detection of 172 nM. The NG sensors were applied in a small pilot test on five airway samples from five CF patients. The NG sensor was capable of identifying P. aeruginosa in the airway samples in 60 s without any sample pretreatment.Graphical abstractImage 1
       
  • Determination of monoamine neurotransmitters and metabolites by
           high-performance liquid chromatography based on Ag(III) complex
           chemiluminescence detection
    • Abstract: Publication date: Available online 21 January 2020Source: Analytical BiochemistryAuthor(s): Li Ma, Tangjuan Zhao, PingPing Zhang, Mengying Liu, Hongmei Shi, Weijun KangA novel, simple and efficient chemiluminescence system has been developed for the determination of monoamine neurotransmitters and metabolites. By using the Ag (III)-luminol chemiluminescence system as a detector, a high performance liquid chromatography chemiluminescence method (HPLC-CL) was established and used to detect seven monoamine neurotransmitters. Under the optimized conditions, the detection limits (3S/N) of epinephrine (E), levodopa (l-DOPA), dopamine (DA), serotonin (5-HT), 3-methoxy-4-hydroxyphenylglycol (MHPG), 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxypentylacetic acid (5-HIAA) were 20.0 μg dm−3,15.0 μg dm−3, 15.0 μg dm−3, 8.0 μg dm−3, 2.0 μg dm−3, 2.0 μg dm−3 and 3.0 μg dm−3, respectively. Moreover, they were well within the linear range of 50–1000 μg dm−3, 50–1000 μg dm−3, 50–1000 μg dm−3, 25–1000 μg dm−3, 5–25 μg dm−3, 5–25 μg dm−3 and 10–30 μg dm−3, respectively. The average recovery varied between 84.82% and 110.4%. The method has the attributes of simplicity, high sensitivity, and high efficiency. The sensitization and inhibition mechanisms for luminol-[Ag(HIO6)2]5–- analytes CL system were proposed by CL spectra and free-radical capture experiment.Graphical abstractImage 1
       
  • Sensitive and label-free liquid crystal-based optical sensor for the
           detection of malathion
    • Abstract: Publication date: Available online 21 January 2020Source: Analytical BiochemistryAuthor(s): Pham Thi Kim Hong, Chang-Hyun JangIn this paper, we report the development of a rapid and simple, liquid crystal (LC)-based aptasensor that enables the detection of malathion (MA) using the orientation properties of liquid crystals. This sensor is composed of aptamers immobilized on a surface decorated with a self-assembled monolayer of (3-glycidyloxypropyl)trimethoxysilane (GOPS) and dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride (DMOAP). When MA interacts with the immobilized aptamers, an orientational change in the LCs, from homeotropic to random, is induced. This orientational change generates visible optical responses observed as shifts from dark to bright images under a polarized optical microscope (POM). This sensing system has a linear detection range from 0.8 to 50 pM, with a correlation coefficient of 0.9922, and a limit of detection (LOD) of 2.5 pM (≈0.826 pg/mL). Our proposed aptasensor has good specificity and sensitivity to MA in tap water and soil. Moreover, this sensor suggests a promising strategy for simple, rapid testing for various insecticide residues.Graphical abstractImage 1
       
  • Primer design strategy for denaturation bubble-mediated strand exchange
           amplification
    • Abstract: Publication date: Available online 21 January 2020Source: Analytical BiochemistryAuthor(s): Jie Deng, Yang Li, Wenqiang Shi, Rui Liu, Cuiping Ma, Chao ShiDenaturation bubble-mediated strand exchange amplification (SEA) is a novel, rapid isothermal nucleic acid amplification has been applied for point-of-care molecular diagnostic in food safety, meat adulteration, forest disease and animal disease. Nevertheless, the absence of specialized strategy for SEA primers design led to long-time of primer screening progress before SEA reaction execution, which would largely increase the time consuming when SEA is utilized for detecting other new targets. In this present work, we investigated the impact of the following primers' attributes on SEA efficiency, including Tm value, 3′ end G/C content, self-complementary and 3′ complementary, according to which we demonstrated that optimal Tm value and reaction temperature were all 61 °C, while 3′-terminal nucleotide should be G/C, as the SEA reaction induced by the primers possessing these attributes exhibited significantly lower threshold time (Tt) value. Moreover, self-complementary and 3′ complementary of primers should be avoided. Besides, we also discussed the consideration priority order of these factors, which was self-complementary and 3′ complementary, Tm value and 3′ end G/C content in turn. Because the SEA primer design strategy is first presented, our work will greatly promote the application of SEA in point-of-care test.Graphical abstractImage 1
       
  • Inspector: A lysine succinylation predictor based on edited
           nearest-neighbor undersampling and adaptive synthetic oversampling
    • Abstract: Publication date: Available online 20 January 2020Source: Analytical BiochemistryAuthor(s): Yan Zhu, Cangzhi Jia, Fuyi Li, Jiangning SongLysine succinylation is an important type of protein post-translational modification and plays a key role in regulating protein function and structural changes. The mechanism and function of succinylation have not been clarified. The key to better understanding the precise mechanism and functional role of succinylation is the identification of lysine succinylation sites. However, conventional experimental methods for succinylation identification are often expensive, time-consuming, and labor-intensive. Therefore, the new development of computational approaches to effectively identify lysine succinylation sites from sequence data is much needed. In this study, we proposed a novel predictor for lysine succinylation identification, Inspector, which was developed by using the random forest algorithm combined with a variety of sequence-based feature-encoding schemes. Edited nearest-neighbor undersampling method and adaptive synthetic oversampling approach were employed to solve dataset imbalance, and a two-step feature-selection strategy was applied to optimize the feature set for training the accuracy of the prediction model. Empirical studies on performance comparison with existing tools showed that Inspector was able to achieve competitive predictive performance for distinguishing lysine succinylation sites.Graphical abstractImage 1
       
  • Development of a lateral flow aptamer assay strip for facile
           identification of theranostic exosomes isolated from human lung carcinoma
           cells
    • Abstract: Publication date: Available online 20 January 2020Source: Analytical BiochemistryAuthor(s): Qiang Yu, Qu Zhao, Sai Wang, Shuai Zhao, Shan Zhang, Yingai Yin, Yiyang DongExosomes are Extracellular Vesicles (EV) that own unique structural features and functions and have gradually become the hot research spot in recent years. The tumor-derived exosomes contain various types of useful biological information, and medical identification of exosomes relied on the specific characterization of membrane surface proteins. In this study, in order to rapidly identify non-small cell lung cancer (NSCLC)-derived exosomes, based on an aptamer against CD63 protein on exosome membrane, a low cost lateral flow aptamer assay (LFAA) test strip using nanogold particles as visualization probes was successfully developed for facile identification of A549 exosomes isolated from human lung carcinoma cells diluted from 6.4 × 109 particles/mL herein.Graphical abstractImage 1
       
  • Biotin oligonucleotide labeling reactions: A method to assess their
           effectiveness and reproducibility
    • Abstract: Publication date: Available online 18 January 2020Source: Analytical BiochemistryAuthor(s): Aldo Di Vito, Erika Reitano, Luisa Poggi, Margherita IaboniThe strong molecular interaction between biotin and streptavidin is widely used in the growing field of nucleic acid nanotechnology. Several biotin labeled oligonucleotide tools have been developed for the detection of biological molecules as well as for protein purification. For these reasons, biotinylation can be considered one of the main chemical reactions for nucleic acid labeling. However, despite its widespread application and the presence on the market of many reagents for the conjugation of biotin to oligonucleotides, it is not yet available a cheap, easy and sensitive system able to assess the effectiveness and reproducibility of this reaction. Here, we present an accurate and reliable method to achieve a qualitative and quantitative analysis of oligonucleotide biotinylation. The protocol employs basic laboratory instruments and standard software for molecular biology applications and does not require advanced expertise for its execution. Most importantly, our method is independent from complex kinetic equilibrium parameters and shows a limit of detection more than one order of magnitude lower than the current fluorometric gold standard assay. Therefore, this method could become a standard, inexpensive and routinely used quality test for post-synthesis evaluation of biotin conjugation reactions.Graphical abstractImage 1
       
  • A proposed sample handling of ovine cotyledon for proteomic studies
    • Abstract: Publication date: Available online 16 January 2020Source: Analytical BiochemistryAuthor(s): M.A. El-Samahy, Xiaolei Yao, Guomin Zhang, Yanli Zhang, Feng WangOvine trophoblast is a suitable material for placental studies. However, a universal protocol for handling ruminant cotyledon samples has still not been reported. Considering the villous structure of ovine cotyledon, we suggest procedures to prepare cotyledons with limited inherent contamination, using semi-dry conditions to avoid freezing damage and sample errors. The cytosolic water-soluble proteins were physically extracted from the frozen cotyledons. High homogeneity was demonstrated between the replicates of both tissue and extract samples. Importantly, the chemical lysis of placental crude extracts was necessary for protein separation and immunoreaction. The integrity of stored tissues was histologically validated using a formalin-fixed paraffin-embedded technique. Using label-free proteomics, we detected 388 Ovis protein-groups in at least two of three biological replicates of either the tissue or extract. Although the water-soluble proteins were dominated by hemoglobin subunits, ten proteins were identified exclusively in all extract replicates. The physical extraction selectively reduced the membrane, extracellular matrix, and cytoskeleton proteins. The hydrolase enzymes, in the extract, hindered the identification of some specific proteins, such as histone H2A. In summary, the proposed workflow may guide further proteomic investigations of ovine cotyledon biology. Furthermore, our proteomic data have inferred some potential mechanisms of ovine trophoblast at parturition.Graphical abstractImage 1
       
  • Microbial transglutaminase: A biotechnological tool to manage gluten
           intolerance
    • Abstract: Publication date: Available online 15 January 2020Source: Analytical BiochemistryAuthor(s): Diomira Luongo, Francesco Maurano, Paolo Bergamo, Mauro RossiAbstractCeliac disease (CD) is a chronic immune-mediated disease in which gluten ingestion leads to damage of the small intestinal mucosa in genetically susceptible individuals. The enteropathy is mainly induced by the production of IFN-γ from intestinal CD4+T cells that recognise gliadin peptides following deamidation by tissue transglutaminase. The only available therapy is a strict, lifelong gluten-free diet (GFD). This diet is strongly demanding for patients, which justifies the search for alternative strategies. The enzyme approach is one promising strategy to address this issue. In particular, transamidation of wheat gliadin by microbial transglutaminase (mTG) was fully effective at inhibiting gliadin-specific IFN-γ secretion in intestinal T cells from CD patients. Furthermore, transamidated gliadin induced higher levels of the anti-inflammatory IL-10 than native gliadin in different in vitro models. These data suggest that a more balanced immune response could be induced by mTG-treated gliadin in the small intestine of celiac patients. Furthermore, the highlighted biological property of mTG-treated gliadin could be exploited to induce tolerance to native gliadin in at-risk individuals.
       
  • A cell-based assay system for activators of the environmental cell stress
           response
    • Abstract: Publication date: Available online 13 January 2020Source: Analytical BiochemistryAuthor(s): Jennifer A. Harbottle, Linda Petrie, Madeleine Ruhe, Wael E. Houssen, Marcel Jaspars, Andreas F. KolbImproved health span and lifespan extension in a wide phylogenetic range of species is associated with the induction of the environmental cell stress response through a signalling pathway regulated by the transcription factor Nrf2. Phytochemicals which stimulate this response may form part of therapeutic interventions which stimulate endogenous cytoprotective mechanisms, thereby delaying the onset of age-related diseases and promoting healthy ageing in humans. In order to identify compounds that activate the Nrf2 pathway, a cell-based reporter system was established in HepG2 cells using a luciferase reporter gene under the control of the Nqo1 promoter. Sulforaphane, an isothiocyanate derived from cruciferous vegetables and a known activator of the Nrf2 pathway, was used to validate the reporter system. The transfected cell line HepG2 C1 was subsequently used to screen natural product libraries. Five compounds were identified as activating the bioluminescent reporter by greater than 5-fold. The two most potent compounds, MBC20 and MBC37, were further characterised and shown to stimulate endogenous cytoprotective gene and protein expression. The bioluminescent reporter system will allow rapid, in vitro identification of novel compounds that have the potential to improve health span through activation of the environmental stress response.Graphical abstractImage 1
       
  • Principal component analysis of MALDI-TOF MS of whole-cell foodborne
           pathogenic bacteria
    • Abstract: Publication date: Available online 11 January 2020Source: Analytical BiochemistryAuthor(s): Wenjing Yan, Jing Qian, Yongjie Ge, Keping Ye, Cunshan Zhou, Houseng ZhangThe rapid and accurate identification of foodborne pathogenic bacteria is of great importance for human health. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) can be used to rapidly and sensitively identify microorganisms but is limited by the expensive protein databases available. In this study, we established a whole-cell method for the identification of foodborne pathogenic bacteria, using MALDI-TOF MS and principal component analysis (PCA), which did not use protein extractions or expensive protein databases. Thirty strains comprising six common foodborne pathogenic bacteria, namely, Shigella flexneri, Escherichia coli, Staphylococcus aureus, Salmonella enteritidis, Pseudomonas aeruginosa, and Listeria monocytogenes were analyzed using MALDI-TOF MS. The culture time, matrix, and spotting method were optimized based on peak intensity and deviation. A PCA was performed to analyze the mass spectrometry results of six samples and proved capable of identifying significant changes in those samples. It was found that directly applying MALDI-TOF MS analysis to whole-cell bacteria, without protein extraction, exhibited rich peak contents and a high level of reproducibility. MALDI-TOF MS combined with PCA is a promising method of rapidly identifying pathogens in food products.Graphical abstractImage 1
       
  • Nano differential scanning fluorimetry for comparability studies of
           therapeutic proteins
    • Abstract: Publication date: Available online 11 January 2020Source: Analytical BiochemistryAuthor(s): Jie Wen, Harrison Lord, Nicholas Knutson, Mats WikströmDifferential scanning calorimetry (DSC) has been extensively used in the biopharmaceutical industry to characterize protein thermal stability and domain folding integrity. Recently, nano differential scanning fluorimetry (nanoDSF) has emerged as a powerful tool for thermal stability analysis and studies of protein domain unfolding. Due to increased interests in the qualification of characterization methods, we are in this study presenting the qualification results for the comparability studies of thermal stability analysis using nanoDSF. The results show that nanoDSF is able to detect thermal transition signals for mAbs, BiTE® molecules, and cytokines at a wide concentration range with high precision, clearly indicating that nanoDSF is suitable for characterization including comparability studies of therapeutic proteins. Compared to the current recognized industry standard DSC, the nanoDSF method enables thermal stability analysis over a much wider concentration range, consumes considerably less materials, and provides significantly higher throughput.Graphical abstractImage 1
       
  • The impact of different human IgG capture molecules on the kinetic
           analysis of antibody-antigen interaction
    • Abstract: Publication date: Available online 10 January 2020Source: Analytical BiochemistryAuthor(s): Vishal Kamat, Ashique Rafique, Tammy Huang, Olav Olsen, William OlsonSurface plasmon resonance (SPR) is a well-established method to characterize biomolecular interactions and is widely used in drug discovery and development. Here, we demonstrate that capture surfaces profoundly impact the binding kinetic parameters that are measured for antibody-antigen interactions. Six unique antibody-antigen interactions were characterized using eight different anti-human IgG capture surfaces. The antigen binding affinities for six different human monoclonal antibodies (hmAbs) captured using three different goat anti-human Fc (AHC) polyclonal antibody (pAb) surfaces were in reasonable agreement (3-7-fold weaker) with those measured by kinetic exclusion assay (KinExA). In contrast, up to 81, 32, 489, 2826, and 219-fold weaker antigen binding affinities were measured using mouse AHC mAb, Protein G, Protein A, Protein A/G, and Protein L surfaces, respectively. Protein A, Protein A/G and Protein G interacted with the Fab of hmAbs, possibly affecting antigen binding to hmAbs captured over these surfaces. Additional studies revealed that mouse AHC mAb binds hmAbs with a weak affinity (5.5–36.3 nM) and t½ values of 1.4–3.3min, compared to the sub-nanomolar affinities of the goat AHC pAbs. These results emphasize the value of measuring binding kinetics of the capture molecule before immobilizing them onto the sensor surface to perform capture kinetic assays on label-free biosensors.Graphical abstractImage 1
       
  • A sensitive and reversible staining of proteins on blot membranes
    • Abstract: Publication date: Available online 9 January 2020Source: Analytical BiochemistryAuthor(s): Jun-Ling Wang, Zhao Li, Mei-Qi Li, Wei-Guang Chen, Chao-Jin XuA modified, sensitive and reversible method for protein staining on nitrocellulose (NC) and polyvinylidine fluoride (PVDF) membranes was developed in Western blotting. The method employed Congo red staining to visualize proteins on different blot membranes. Staining of proteins with Congo red dye is more faster procedures. According to the experimental results, approximate 20 ng proteins could be detected in 3 min in room temperature. The staining on the proteins is easily reversible with Congo red destaining solution for NC and PVDF membranes, so that the blot membranes can be reused for Western blotting. In addition, we confirmed that the staining method is fully compatible with Western blot detection. NC and PVDF membranes treatment with Congo red staining does not interfere with conventional chemiluminescent substrates of peroxidase. As compared to MemCode reversible protein stain kits from Pirece Biotechnology, the staining technique is more sensitive, lower of cost, convenient and not adversely affecting subsequent Western blotting results. On the other hand, the stain is more sensitive than the Ponceau S staining. Therefore, Congo red staining is a promising and ideal alternative for current protein stain. Besides, the binding modes of Congo red or Ponceau S stain were investigated using various 2D and 3D molecular docking and demonstrated potential molecular basis for sensitivity of Congo red staining are higher than Ponceau S.Graphical abstractImage 1
       
  • Interaction between tissue transglutaminase and amyloid-beta:
           Protein-protein binding versus enzymatic crosslinking
    • Abstract: Publication date: Available online 8 January 2020Source: Analytical BiochemistryAuthor(s): Micha M.M. Wilhelmus, Cornelis A. Jongenelen, John G.J.M. Bol, Benjamin DrukarchSelf-interaction, chaperone binding and posttranslational modification of amyloid-beta (Aβ) is essential in the initiation and propagation of Aβ aggregation. Aggregation results in insoluble Aβ deposits characteristic of Alzheimer's disease (AD) brain lesions, i.e. senile plaques and cerebral amyloid angiopathy. Tissue transglutaminase (tTG) is a calcium-dependent enzyme that catalyzes posttranslational modifications including the formation of covalent ε-(γ-glutamyl)lysine isopeptide bonds (molecular crosslinks), and colocalizes with Aβ deposits in AD. Two independent groups recently found that apart from the induction of Aβ oligomerization, the blood-derived transglutaminase member FXIIIa forms stable protein-protein complexes with Aβ independent of the transamidation reaction. Here, we investigated whether also tTG forms rigid protein complexes with Aβ in the absence of catalytic activation. We found that both Aβ1-40 and Aβ1-42 are substrates for tTG-catalyzed crosslinking. In addition, in the absence of calcium or the presence of a peptidergic inhibitor of tTG, stable tTG-Aβ1-40 complexes were found. Interestingly, the stable complexes between tTG and Aβ1-40, were only found at ‘physiological’ concentrations of Aβ1-40. Together, our data suggest that depending on the Aβ species at hand, and on the concentration of Aβ, rigid protein-complexes are formed between tTG and Aβ1-40 without the involvement of the crosslinking reaction.Graphical abstractImage 1
       
  • Development and fit-for-purpose verification of an LC-MS method for
           quantitation of hemagglutinin and neuraminidase proteins in influenza
           virus-like particle vaccine candidates
    • Abstract: Publication date: Available online 8 January 2020Source: Analytical BiochemistryAuthor(s): Jingzhong Guo, Yali Lu, Yun Zhang, Sheila Mugabe, Ziping Wei, Oleg V. BorisovRecombinant influenza Virus-Like Particle (VLP) vaccines are promising vaccine candidates to prevent influenza, contain two major viral antigenic glycoproteins, Hemagglutinin (HA) and Neuraminidase (NA), on the surface of recombinant VLPs. Accurate quantitation of the mass of these antigenic proteins is important to ensure the product quality and proper dosing. Currently, Single Radial Immunodiffusion (SRID) is a recognized assay for determination of the HA immuno-reactive concentration (potency) in vaccine products, based on immuno-reactivity of HA with strain-specific antisera. The SRID assay, however, requires availability of strain-specific and properly calibrated reagents, which can be time-consuming to generate and calibrate. In addition, the assay is not suitable for quantitation of low abundant proteins, such as NA. In order to accelerate the overall production cycle, we have developed and optimized a high-resolution (HR) LC-MS method for absolute quantitation of both HA and NA protein concentrations in influenza VLP vaccine candidates. In this work, we present the method development, optimization and verification of its suitability for the intended purpose, as a prerequisite for its potential application in Quality Control, by assessing specificity, precision and accuracy, detection characteristics, and dynamic linear range. The method can be also used for other HA/NA containing preparations including in-process samples, purified proteins, whole virus preparations, nano-particle and egg-based vaccine preparations, or for calibration of SRID reference antigens.Graphical abstractImage 1
       
  • Development of a revised ICC-qPCR method used for Pseudorabies virus
           inactivation validation study of biologically sourced materials
    • Abstract: Publication date: Available online 7 January 2020Source: Analytical BiochemistryAuthor(s): Yu Zhang, Le Zhang, Xiaojie Duan, Shuxin Qu, Liming XuTo develop a precise and convenient method to evaluate the virus transmission risk of biologically sourced materials, an integrated cell culture-qPCR (ICC-qPCR) method for Pseudorabies virus (PRV) was established and revised for applications to this new field. The optimized post-infection period was found at 12-hr to achieve a reasonable detection limit (−0.25 Log10TCID50/100 μL, Logs) and a quantitative range (0.75–3.75 Logs). The results of mimic samples suggested that three 10-fold dilutions at the time of virus inoculation combined with three washes after virus absorption, and the sets of non-amplified samples as controls could efficiently eliminate the false positive signals caused by high levels of noninfectious viruses. The virus inactivation validation studies of acellular porcine corneas suggested that the logs inactivation of PRV at 12 kGy irradiation dose obtained by general ICC-qPCR, revised ICC-qPCR and cell culture were 2.49, 4.85 and 5.08, respectively. At 25 kGy, those were 2.31, 4.85 and 5.08, respectively. The results obtained by the revised ICC-qPCR were consistent with cell culture and more precise than general ICC-qPCR. Therefore, the revised ICC-qPCR proposed in this study has an application prospect in the PRV inactivation validation studies of biologically sourced materials.Graphical abstractImage 1
       
  • Measuring the affinity of protein-protein interactions on a
           single-molecule level by mass photometry
    • Abstract: Publication date: Available online 7 January 2020Source: Analytical BiochemistryAuthor(s): Di Wu, Grzegorz PiszczekMeasurements of biomolecular interactions are crucial to understand the mechanisms of the biological processes they facilitate. Bulk-based methods such as ITC and SPR provide important information on binding affinities, stoichiometry, and kinetics of interactions. However, the ensemble averaging approaches are not able to probe the intrinsic heterogeneity often displayed by biological systems. Interactions that involve cooperativity or result in the formation of multicomponent complexes pose additional experimental challenges. Single-molecule techniques have previously been applied to solve these problems. However, single-molecule experiments are often technically demanding and require labeling or immobilization of the molecules under study. A recently developed single-molecule method, mass photometry (MP), overcomes these limitations. Here we applied MP to measure the affinities of biomolecular interactions. We have demonstrated how MP allows the user to study multivalent complexes and quantify the affinities of different binding sites in a single measurement. Results obtained from this single-molecule technique have been validated by ITC and BLI. The quality and information content of the MP data, combined with simple and fast measurements and low sample consumption makes MP a new preferred method for measuring strong protein-protein interactions.Graphical abstractImage 1
       
  • A sandwich-type electrochemical immunosensor for detecting CEA based on
           CeO2-MoS2 absorbed Pb2+
    • Abstract: Publication date: Available online 3 January 2020Source: Analytical BiochemistryAuthor(s): Wenjun Li, Xiuwen Qiao, Chenglin Hong, Chaoyun Ma, Yiju SongA sandwich-type immunosensor for detecting the concentration of carcinoembryonic antigen (CEA) was prepared. In this work, gold nanoparticles (Au NPs) were used as platform to attach more primary antibody (Ab1) due to excellent electrical conductivity and good biocompatibility. Molybdenum disulfide-Cerium oxide (CeO2-MoS2) nanohybrid was used as a carrier to absorb lead ions (Pb2+) and the second antibody (Ab2). CeO2-MoS2-Pb2+-Ab2 was used as a nanoprobe to detect CEA antigen. Under optimal conditions, square wave voltammetry (SWV) successfully displayed the electrical signal of Pb2+. The designed electrochemical immunosensor has excellent analytical performance. In addition, the detection range was 0.001–80 ng/mL and the minimum detection limit was 0.3 pg/mL (S/N = 3), which had good selectivity and stability. Finally, the proposed immunosensor successfully detected the concentration of CEA in the serum of the sample, which provided a feasible method for CEA testing.Graphical abstractImage 1
       
  • Robotic automation of a UHPLC/MS-MS method profiling one-carbon
           metabolites, amino acids, and precursors in plasma
    • Abstract: Publication date: Available online 3 January 2020Source: Analytical BiochemistryAuthor(s): Stephanie Andraos, Michael Goy, Benjamin B. Albert, Martin Kussmann, Eric B. Thorstensen, Justin M. O'SullivanAmino acids (AAs) and one-carbon (1-C) metabolism compounds are involved in a range of key metabolic pathways, and mediate numerous health and disease processes in the human body. Previous assays have quantified a limited selection of these compounds and typically require extensive manual handling. Here, we describe the robotic automation of an analytical method for the simultaneous quantification of 37 1-C metabolites, amino acids, and precursors using reversed-phase ultra-high-pressure liquid chromatography coupled with tandem mass spectrometry (UHPLC/MS-MS). Compound extraction from human plasma was tested manually before being robotically automated. The final automated analytical panel was validated on human plasma samples. Our automated and multiplexed method holds promise for application to large cohort studies.Graphical abstractImage 1
       
  • Application of aptamers as molecular recognition elements in lateral flow
           assays for analytical applications
    • Abstract: Publication date: Available online 3 January 2020Source: Analytical BiochemistryAuthor(s): Ruth Reid, Bandhan Chaterjee, Soonjyoti Das, Sourav Ghosh, Tarun Kumar SharmaAbstractOwing to their ease in operation and fast turnaround time, lateral flow assays (LFAs) are increasingly being used as point-of-care diagnostic tests for a variety of analytes. In a majority of these LFAs, antibodies are used as a molecular recognition element. Antibodies have a number of limitations such as high batch-to-batch variation, poor stability, long development time, difficulty in functionalization and need for ethical approval and cold chain. All these factors pose a great challenge to scale up the antibody-based tests. In recent years, the advent of aptamer technology has made a paradigm shift in the point-of-care diagnostics owing to the various advantages of aptamers over antibodies that favour their adaptability on a variety of sensing platforms including the lateral flow. In this review, we have highlighted the advantages of aptamers over antibodies, suitability of aptamers for lateral flow platforms, different types of aptamer-based LFAs and various labels for aptamer-based LFAs. We have also provided a summary of the applications of aptamer technology in LFAs for analytical applications.
       
  • Phosphorescent iridium(III) complex for efficient sensing of hypochlorite
           and imaging in living cells
    • Abstract: Publication date: Available online 31 December 2019Source: Analytical BiochemistryAuthor(s): Jinsheng Liu, Mingqin Shangguan, Xiaoyang Zeng, Yingxiong Guo, Tao Wang, Linxi HouIn consideration of the strong oxidizing power of hypochlorite (ClO−), which could cleave CN moiety, a cyclometalated iridium (III) complex (Ir-Ts) modified hydrazide group as the response unit was synthesized to sensitively and selectively detect ClO− under neutral condition. Upon addition of ClO−, a 21-fold emission enhancement at 574 nm was observed and phosphorescent product was formed due to the cleavage of CN moiety. The probe Ir-Ts displayed rapid response (
       
  • Design of customizable long linear DNA substrates with controlled end
           modifications for single-molecule studies
    • Abstract: Publication date: Available online 20 December 2019Source: Analytical BiochemistryAuthor(s): Stefan H. Mueller, Lisanne M. Spenkelink, Antoine M. van Oijen, Jacob S. LewisMany strategies have been developed to manipulate DNA molecules and investigate protein–DNA interactions with single-molecule resolution. Often, these require long DNA molecules with a length of several 10s of kb that are chemically modified at specific regions. This need has traditionally been met by commercially available DNA from bacteriophage λ. However, λ DNA does not allow for the generation of highly customizable substrates in a straightforward manner, an important factor when developing assays to study complex biochemical reactions. Here we present a generalizable method for the design and production of very long chemically modified DNA substrates derived from a single plasmid. We show the versatility of this design by demonstrating its application in studying DNA replication in vitro. We anticipate this strategy will be broadly useful in producing a range of long chemically modified DNA molecules required for a diverse range of single-molecule approaches.Graphical abstractImage 1
       
  • Rapid detection of DPP-IV activity in porcine serum: A fluorospectrometric
           assay
    • Abstract: Publication date: Available online 20 December 2019Source: Analytical BiochemistryAuthor(s): K. Divya, H.K. Vivek, B.S. Priya, S. Nanjunda SwamyDipeptidyl peptidase IV (DPP-IV) is an aminopeptidase that cleaves the N-terminal dipeptide from peptides bearing proline or alanine residues. Currently, DPP-IV activity is quantified by spectrophotometric or fluorometric methods, which employ Gly-Pro-pNA and Gly-Pro-AMC respectively, as substrate. However, these methods require high enzyme and substrate concentrations. In this study, we adapted the DPP-IV fluorospectrometric assay using NanoDrop 3300, which requires only nanogram levels of the enzyme (30 ng crude DPP-IV) and considerably low substrate concentrations (100 μM). Fluorescence measurement required a reaction mixture of only 2 μL, thus eliminating the need for microtiter plates or cuvettes.We employed this assay to demonstrate DPP-IV activity in porcine serum for the first time. The enzymatic activity peaked at pH 8.0 in porcine (84 nM/min), human (87 nM/min) and bovine (89.1 nM/min) sera, with the optimum temperature of 37 °C. The enzyme showed maximum activity upon incubation for 40 min at 37 °C. In contrast, activity in the porcine serum was the highest after incubation for 30 min at the same optimized parameters. The IC50 values of diprotin A against DPP-IV from human, porcine, and bovine sera were 7.83, 8.62, 9.17 μM, respectively. The present assay procedure is a convenient, sensitive, accurate and high-throughput method suitable for primary screening of DPP-IV inhibitors.Graphical abstractFluoro spectrometric assay of DPP-IV by NanoDrop.Image 1
       
  • An inverse cell culture model for floating plastic particles
    • Abstract: Publication date: Available online 14 December 2019Source: Analytical BiochemistryAuthor(s): Valerie Stock, Linda Böhmert, Merve Hilal Dönmez, Alfonso Lampen, Holger SiegAbstractPlastic waste has become a major environmental problem. An increasing number of studies investigate microplastic particles with regard to their uptake and effects in cell culture systems. Individual plastic materials vary in their molecular structure, composition, size distribution, material density, and may also differ with respect to their toxicological effects. Plastic particles with lower densities than the cell culture medium, for example polyethylene (PE), pose a particular problem for in vitro assays as they float up during the incubation and thus do not contact the cells located on the bottom of the culture dish. We thus developed a practical and easy-to-use in vitro inverse cell culture model for investigating cellular effects of floating plastic particles. Cytotoxicity tests with floating PE particles were performed to demonstrate the utility of the inverted cell model. PE particles incubated in overhead culture were cytotoxic to HepG2 cells, while under the same cultivation conditions, except for inversion, no cytotoxicity occurred. These positive results demonstrate that inverted cell culture was required to detect the effects of PE particles and underlines the necessity to adapt cell culture conditions to the physicochemical properties of particles in order to obtain a more accurate estimate of the effects of floating particles on cells.
       
  • Rapid isolation of bacteria-specific aptamers with a non-SELEX-based
           method
    • Abstract: Publication date: Available online 12 December 2019Source: Analytical BiochemistryAuthor(s): Hye Ri Kim, Min Yong Song, Byoung Chan KimAbstractUsually, isolation of bacteria-specific aptamers by SELEX is a time-consuming process due to the required repeated rounds of binding, separation, and amplification of the probes to target bacteria. Here, we show that it is possible to isolate bacteria-specific DNA aptamers omitting the repeated rounds of binding incubation, separation, and amplification that are indispensable for SELEX. The serial removal of unbound DNAs to the bacterial cells from an initial mixture of bacteria and DNA libraries through serial centrifugation, one-time separation, and further one-time amplification of DNA bound to the target bacterial cells applied in this non-SELEX-based method allows successful aptamer isolation.
       
  • Vinblastine production by the endophytic fungus Curvularia verruculosa and
           their in vitro cytotoxicity
    • Abstract: Publication date: Available online 30 November 2019Source: Analytical BiochemistryAuthor(s): Ramalingam Parthasarathy, Rajasree Shanmuganathan, Arivalagan PugazhendhiThe current study was to isolate endophytic fungi producing high yields of indole alkaloids such as vinblastine analogous to their host Cathranthus roseus. Endophytic fungi were isolated from the leaves of C. roseus, identified as Curvularia verruculosa by molecular techniques, the sequence was deposited in NCBI (MK995628). Vinblastine producing endophytic fungus was grown in 1L Vinca medium for 21 days. The extract was examined for vinblastine by chromatographic techniques. TLC plates showed purple colour spot co migrated with authentic vinblastine and Rf was calculated by HPTLC (Vin 1 vinblastine −0.75; authentic vinblastine-0.78), these results confirmed vinblastine presence in the Vin1 extract. Further, the TLC purified fungal extract was examined by LC-MS, which revealed the exact mass of vinblastine ([M + H]+ m/z 811.51). The most important of the study is high yield production of vinblastine; hence, the extract analysed by HPLC revealed 182 μg/L vinblastine. The TLC purified fungal vinblastine was analysed for the cytotoxicity effect on HeLa cell line and it depicted a higher activity with IC50-8.5 μg/mL and apoptotic morphological changes were analysed. All the results revealed that the endophytic fungus Curvularia verruculosa produced vinblastine and for the first time in a surplus amount compared to other fungi.Graphical abstractImage 1
       
  • Sensitivity of the polyDetect computational pipeline for phylogenetic
           analyses
    • Abstract: Publication date: Available online 30 November 2019Source: Analytical BiochemistryAuthor(s): Jessica M. Storer, Jerilyn A. Walker, Vallmer E. Jordan, Mark A. BatzerAlu elements are powerful phylogenetic markers. The combination of a recently-developed computational pipeline, polyDetect, with high copy number Alu insertions has previously been utilized to help resolve the Papio baboon phylogeny with high statistical support. Here, the polyDetect method was applied to the highly contentious Cebidae phylogeny within New World monkeys (NWM). The polyDetect method relies on conserved homology/identity of short read sequence data among the species being compared to accurately map predicted shared Alu insertions to each unique flanking sequence. The results of this comprehensive assessment indicate that there were insufficient sequence homology/identity stretches in non-repeated DNA sequences among the four Cebidae genera analyzed in this study to make this strategy phylogenetically viable. The ∼20 million years of evolutionary divergence of the Cebidae genera has resulted in random sequence decay within the short read data, obscuring potentially orthologous elements in the species tested. These analyses suggest that the polyDetect pipeline is best suited to resolving phylogenies of more recently diverged lineages when high-quality assembled genomes are not available for the taxa of interest.Graphical abstractImage 1
       
 
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