Antibodies Targeting Closely Adjacent or Minimally Overlapping Epitopes Can Displace One Another
العنوان: | Antibodies Targeting Closely Adjacent or Minimally Overlapping Epitopes Can Displace One Another |
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المؤلفون: | Donna Marie Stone, Andy Yik Yeung, Yasmina Noubia Abdiche, Adam Miles, Pavel Strop, Winse Morishige, Andrea Rossi, Irene Ni |
المصدر: | PLoS ONE, Vol 12, Iss 1, p e0169535 (2017) PLoS ONE |
بيانات النشر: | Public Library of Science (PLoS), 2017. |
سنة النشر: | 2017 |
مصطلحات موضوعية: | Models, Molecular, 0301 basic medicine, Protein Conformation, Physiology, Antibody Affinity, lcsh:Medicine, Biosensing Techniques, Biochemistry, Poultry, Epitope, Epitopes, Binding Analysis, 0302 clinical medicine, Protein structure, Immune Physiology, Medicine and Health Sciences, Cluster Analysis, Gamefowl, lcsh:Science, Immune System Proteins, Multidisciplinary, biology, Chemistry, Antibodies, Monoclonal, food and beverages, Detectors, Agriculture, Hydrogen-Ion Concentration, 030220 oncology & carcinogenesis, Vertebrates, Engineering and Technology, Antibody, Concentration Analysis, Antibody Production, Protein Binding, Research Article, Analyte, Livestock, Immunology, Equipment, Research and Analysis Methods, Antibodies, Birds, 03 medical and health sciences, Antigen, Epitope binning, Humans, Animals, Antigens, Molecular Biology Techniques, Molecular Biology, Chemical Characterization, Gene Mapping, lcsh:R, Organisms, Biology and Life Sciences, Proteins, High-Throughput Screening Assays, Kinetics, Biosensors, 030104 developmental biology, Epitope mapping, Fowl, Amniotes, Immunologic Techniques, biology.protein, Biophysics, lcsh:Q, Chickens, Biosensor, Epitope Mapping |
الوصف: | Here we describe how real-time label-free biosensors can be used to identify antibodies that compete for closely adjacent or minimally overlapping epitopes on their specific antigen via a mechanism of antibody displacement. By kinetically perturbing one another’s binding towards their antigen via the formation of a transient trimolecular complex, antibodies can displace one another in a fully reversible and dose-dependent manner. Displacements can be readily identified when epitope binning assays are performed in a classical sandwich assay format whereby a solution antibody (analyte) is tested for binding to its antigen that is first captured via an immobilized antibody (ligand) because an inverted sandwiching response is observed when an analyte displaces a ligand, signifying the antigen’s unusually rapid dissociation from its ligand. In addition to classifying antibodies within a panel in terms of their ability to block or sandwich pair with one another, displacement provides a hybrid mechanism of competition. Using high-throughput epitope binning studies we demonstrate that displacements can be observed on any target, if the antibody panel contains appropriate epitope diversity. Unidirectional displacements occurring between disparate-affinity antibodies can generate apparent asymmetries in a cross-blocking experiment, confounding their interpretation. However, examining competition across a wide enough concentration range will often reveal that these displacements are reversible. Displacement provides a gentle and efficient way of eluting antigen from an otherwise high affinity binding partner which can be leveraged in designing reagents or therapeutic antibodies with unique properties. |
اللغة: | English |
تدمد: | 1932-6203 |
URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::891aff11f83be4d19e4eddaeb2f5b35d http://europepmc.org/articles/PMC5218414?pdf=render |
حقوق: | OPEN |
رقم الأكسشن: | edsair.doi.dedup.....891aff11f83be4d19e4eddaeb2f5b35d |
قاعدة البيانات: | OpenAIRE |
تدمد: | 19326203 |
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