Recurrent somatic mutations in mouse antibodies to p-azophenylarsonate increase affinity for hapten.
Two mouse mAb specific for the hapten p-azophenylarsonate and encoded by the same combination of germ-line V, D, and J genes differ 200-fold in affinity for hapten. We determined the amino acid sequences of the V regions of the high affinity antibody and compared them to the published sequences of the low affinity antibody which is not somatically mutated. Of 19 amino acid substitutions, two, Ile57 and Thr58 in the H chain, also occur, either alone or together, in other somatically mutated antibodies specific for p-azophenylarsonate; these antibodies have been independently isolated. Introduction of either one of these mutations alone into the low affinity antibody by oligonucleotide-directed mutagenesis increased the antibody affinity for hapten three- to fourfold, whereas introduction of both mutations together conferred an eightfold increase in affinity. These results support the hypothesis that somatic mutations are selected on the basis of the affinity for antigen that they confer, and suggest that even relatively small increases in affinity may be selected, probably in a sequential manner.
Sharon
J
J
Department of Pathology, Hubert H. Humphrey Cancer Research Center, Boston, MA 02118.
Gefter
M L
ML
Wysocki
L J
LJ
Margolies
M N
MN
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification*; D000915:Antibody Affinity; D001391:Azo Compounds / Q000276:immunology*; D001666:Binding Sites, Antibody; D006241:Haptens / Q000276:immunology*; D006822:Hybrid Cells / Q000032:analysis; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D012091:Repetitive Sequences, Nucleic Acid; D010132:p-Azobenzenearsonate / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001391:Azo Compounds; D006241:Haptens; D007135:Immunoglobulin Variable Region; D010132:p-Azobenzenearsonate