The remarkable flexibility of the human antibody repertoire; isolation of over one thousand different antibodies to a single protein, BLyS.
It is well established that the humoral immune response can generate antibodies to many different antigens. The antibody diversity required to achieve this is believed to be substantial. However, the extent to which the immune repertoire can generate structural diversity against a single target antigen has never been addressed. Here, we have used phage display to demonstrate the extraordinary capacity of the human antibody repertoire. Over 1000 antibodies, all different in amino acid sequence, were generated to a single protein, B-lymphocyte stimulator (BLyS protein). This is a highly diverse panel of antibodies as exemplified by the extensive heavy and light chain germline usage: 42/49 functional heavy chain germlines and 19/33 V(lambda) and 13/35 V(kappa) light chain germlines were all represented in the panel of antibodies. Moreover, a high level of sequence diversity was observed in the V(H) CDR3 domains of these antibodies, with 568 different amino acid sequences identified. Thus we have demonstrated that specific recognition of a single antigen can be achieved from many different VDJ combinations, illustrating the remarkable problem-solving ability of the human immune repertoire. When studied in a biochemical assay, around 500 (40%) of these antibodies inhibited the binding of BLyS to its receptors on B-cell lines. The most potent antibodies inhibited BLyS binding with sub-nanomolar IC(50) values and with sub-nanomolar affinities. Such antibodies provide excellent choices as candidates for the treatment of BLyS-associated autoimmune diseases.
Edwards
Bryan M
BM
Cambridge Antibody Technology, Milstein Building, Granta Park, Cambridge CB1 6GH, UK. bryan.edwards@cambridgeantibody.com
Barash
Steven C
SC
Main
Sarah H
SH
Choi
Gil H
GH
Minter
Ralph
R
Ullrich
Stephen
S
Williams
Elizabeth
E
Du Fou
Leila
L
Wilton
Jane
J
Albert
Vivian R
VR
Ruben
Steve M
SM
Vaughan
Tristan J
TJ
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000276:immunology; D053264:B-Cell Activating Factor; D022801:Complementarity Determining Regions; D006801:Humans; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D008565:Membrane Proteins / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D010802:Phylogeny; D017354:Point Mutation; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D000906:Antibodies; D053264:B-Cell Activating Factor; D022801:Complementarity Determining Regions; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D008565:Membrane Proteins; D019151:Peptide Library; C505719:TNFSF13B protein, human; D014409:Tumor Necrosis Factor-alpha