Method for identifying lineage-related antibodies

摘要

In certain embodiments, the method may comprise: a) obtaining the antibody sequences from a population of B cells; b) grouping the antibody sequences to provide a plurality of groups of lineage-related antibodies; c) testing a single antibody from each of the groups in a bioassay and, after the first antibody has been identified, d) testing further antibodies that are in the same group as the first antibody in a second bioassay. In another embodiment, the method may comprise: a) testing a plurality of antibodies obtained from a first portion of an antibody producing organ of an animal; b) obtaining the sequence of a first identified antibody; c) obtaining from a second portion of said antibody producing organ the sequences of further antibodies that are related by lineage to said first antibody; and, c) testing the further antibodies in a second bioassay.

主权项

1. A method for identifying an antibody that binds to an antigen, comprising:
a) obtaining: (i) antibody heavy chain variable domain sequences and (ii) antibody light chain variable domain sequences by sequencing heavy and light chain-encoding cDNAs made from a population of antibody-producing B cells of an animal that has been immunized by the antigen; b) grouping the heavy chain variable domain sequences of step a) into a plurality of heavy chain groups and, independently, grouping the light chain variable domain sequences of step a) into a plurality of light chain groups, wherein:
(i) heavy chain sequences that comprise CDR3 regions that are of the same length and contain up to five amino acid substitutions relative to one another are grouped together; and(ii) light chain sequences that comprise CDR3 regions that are of the same length and contain up to five amino acid substitutions relative to one another are grouped together; c) selecting a heavy chain group and a light chain group from the groups produced by step b); d) pairing a heavy chain variable domain sequence and a light chain variable domain sequence from the groups selected in c); and e) testing an antibody comprising the heavy and light chain sequences paired in d) for binding to the antigen.