Parallel kinetic analysis and affinity determination of hundreds of monoclonal antibodies using the ProteOn XPR36.
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.
Nahshol
Oded
O
Bio-Rad Laboratories, Technion, Haifa 32000, Israel.
Bronner
Vered
V
Notcovich
Ariel
A
Rubrecht
Laetitia
L
Laune
Daniel
D
Bravman
Tsafrir
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology*; D000918:Antibody Specificity; D015374:Biosensing Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas; D018664:Interleukin-12 / Q000276:immunology; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008962:Models, Theoretical
D000911:Antibodies, Monoclonal; D018664:Interleukin-12