Parallel kinetic analysis and affinity determination of hundreds of monoclonal antibodies using the ProteOn XPR36
| Autor: | Laetitia Rubrecht, Tsafrir Bravman, Daniel Laune, Oded Nahshol, Vered Bronner, Ariel G. Notcovich |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
medicine.drug_class
High-throughput screening Kinetic analysis Biophysics Enzyme-Linked Immunosorbent Assay Biosensing Techniques Plasma protein binding Computational biology Monoclonal antibody Biochemistry Mice Antigen Antibody Specificity medicine Animals Humans Molecular Biology Mice Inbred BALB C Hybridomas biology Antibodies Monoclonal Cell Biology Models Theoretical Interleukin-12 Molecular biology Kinetics Monoclonal biology.protein Antibody |
| Zdroj: | Analytical Biochemistry. 383:52-60 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2008.08.017 |
| Popis: | The production of antibodies for diagnostic and therapeutic applications is a major focus for biotechnology and pharmaceutical companies, and it requires the development of fast, high-throughput methodologies for screening and selecting appropriate candidate antibodies for development. These candidates must have very high affinity for the target as well as high specificity and low cross-reactivity. This study demonstrates the use of the ProteOn XPR 36 protein interaction array system for the rapid screening and selection of high-affinity antibodies—“one-shot” kinetics. This approach allows multiple quantitative protein binding analyses in parallel, providing association, dissociation, and affinity constants for several antibodies simultaneously in one experiment. We have used this new methodology to screen hundreds of monoclonal supernatants containing antibodies against two proteins of potential clinical interest: human interleukin 12 (IL-12) and a human hemoglobin (Hb) variant. In fact, approximately 250 supernatants raised against each antigen were screened in approximately 17 h, providing several high-affinity candidate monoclonal antibodies for each of these antigens. |
| Databáze: | OpenAIRE |
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