Monoclonal antibody classification based on epitope-binding using differential antigen disruption.
Currently, classifying a population of specific antigen-reactive monoclonal antibodies (mAbs) according to their epitope-binding properties has been limited to competition assays. Such assays are time consuming, labor intensive and restricted to the number of mAbs in the experiment. To overcome this problem, a differential antigen disruption-based antibody profiling procedure was developed. This procedure rapidly classifies specific antigen-reactive mAbs into epitope-related groups by measuring the binding signal of the antibodies to a set of structurally disrupted antigens and then clustering the antibodies according to the similarity of their binding profiles. The clustering results generated by differential antigen disruption showed a significant concordance with those generated by competition experiments. Therefore, differential antigen disruption method opens an opportunity to assess the entire population of antigen-reactive mAbs according to their epitope-binding properties. In doing so, a set of representative antibodies can be drawn to describe the epitope complexity for systematically exploring their functions.
Shi
Ergang
E
Regeneron Pharmaceuticals Inc., 777 Old Saw Mill River Road, Tarrytown, NY 10591, USA. ergang.shi@regeneron.com <ergang.shi@regeneron.com>
Fury
Wen
W
Li
Wentian
W
Mikulka
Warren
W
Aldrich
Thomas
T
Rafique
Ashique
A
Chen
Gang
G
Hoffenberg
Simon
S
Daly
Thomas J
TJ
Radziejewski
Czeslaw
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000145:classification* / Q000302:isolation & purification; D000918:Antibody Specificity; D000941:Antigens; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D016000:Cluster Analysis; D018604:Epitope Mapping; D000939:Epitopes / Q000032:analysis*; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes