Phage antibody fragments library combining a single human light chain variable region with immune mouse heavy chain variable regions.
We describe the construction of a phage antibody fragments library which combines, in a single cloning step, a synthetic human light chain variable region (V(L)) with a diverse set of heavy chain variable regions, from a mouse immunized with the prostate specific antigen (PSA). Despite V(L) restriction, selection from this library rendered two different single chain Fv antibody fragments, specifically recognizing PSA. The human V(L), used as a general partner for mouse heavy chains, was constructed by linking the germline A27 gene and the J(K)1 minigene segment, both of which are prominently involved in human antibody responses. Our approach offers a fast and simple way to produce half-human molecules, while keeping the advantage of immunizing animals for high affinity antibodies.
Rojas
Gertrudis
G
Recombinant Antibodies Laboratory, Pharmaceuticals Division, P.O. Box 6162, Ave 31 e/158 y 190, Cubanacán, Playa, La Habana 10600, Cuba. gertrudis.rojas@cigb.edu.cu
Almagro
Juan Carlos
JC
Acevedo
Boris
B
Gavilondo
Jorge V
JV
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019151:Peptide Library; D017430:Prostate-Specific Antigen / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D017430:Prostate-Specific Antigen