Methods for producing members of specific binding pairs
First Claim
1. A library of filamentous bacteriophage particles displaying on their surface as a fusion with a gene III coat protein surface component a genetically diverse population of specific binding pair members in functional form comprising a binding domain for complementary binding specific binding pair members, said specific binding pair members encoded by nucleic acid derived from a natural repertoire of nucleic acids encoding said genetically diverse population of specific binding pair members, the particles each containing a phagemid genome which is plasmid nucleic acid containing a single stranded phage replication origin and a nucleotide sequence encoding said fusion, and the particle having a coat partially derived from a helper phage and partly from said fusion.
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Abstract
A member of a specific binding pair (sbp) is identified by expressing DNA encoding a genetically diverse population of such sbp members in recombinant host cells in which the sbp members are displayed in functional form at the surface of a secreted recombinant genetic display package (rgdp) containing DNA encoding the sbp member or a polypeptide component thereof, by virtue of the sbp member or a polypeptide component thereof being expressed as a fusion with a capsid component of the rgdp. The displayed sbps may be selected by affinity with a complementary sbp member, and the DNA recovered from selected rgdps for expression of the selected sbp members. Antibody sbp members may be thus obtained, with the different chains thereof expressed, one fused to the capsid component and the other in free form for association with the fusion partner polypeptide. A phagemid may be used as an expression vector, with said capsid fusion helping to package the phagemid DNA. Using this method libraries of DNA encoding respective chains of such multimeric sbp members may be combined, thereby obtaining a much greater genetic diversity in the sbp members than could easily be obtained by conventional methods.
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Citations
6 Claims
- 1. A library of filamentous bacteriophage particles displaying on their surface as a fusion with a gene III coat protein surface component a genetically diverse population of specific binding pair members in functional form comprising a binding domain for complementary binding specific binding pair members, said specific binding pair members encoded by nucleic acid derived from a natural repertoire of nucleic acids encoding said genetically diverse population of specific binding pair members, the particles each containing a phagemid genome which is plasmid nucleic acid containing a single stranded phage replication origin and a nucleotide sequence encoding said fusion, and the particle having a coat partially derived from a helper phage and partly from said fusion.
Specification