Methods for producing specific binding pairs
AssigneeDyax Corp.
InventorRobert C. Ladner
Life of this patent
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Abstract
Provided are improved methods for providing specific binding pairs (SBPs). The methods enable production of libraries of SBP members using both a large population of one member of the SBPs and a smaller, preselected population of the other member of the SBPs having affinity for a preselected target.
Claims21
A method of producing specific binding pair (SBP) members with affinity for a predetermined target, wherein the SBP comprises a first polypeptide chain and a second polypeptide chain, which method comprises:
- (i) providing host cells that comprise a first population of vectors comprising a population of genetic material encoding one or more of the first polypeptide chains which have been selected to have one or more desirable properties, wherein the first polypeptide chains are secreted from the host cells;
- (ii) infecting the cells with a second population of vectors that comprises a diverse population of genetic material that encodes the second polypeptide chains, wherein the second polypeptide chain is fused to a component of a secreted replicable genetic display package (RGDP) for display of the second polypeptide chains at the surface of RGDPs;
- (iii) expressing the first and second polypeptide chains within the host cells to form a library of SBP members displayed at the surface of the RGDPs, wherein the first and second polypeptide chains are associated at the surface of the RGDPs;
- and
- (iv) selecting SBP members for binding to the predetermined target.
A method of producing specific binding pair (SBP) members with improved affinity for a predetermined target, wherein the SBP comprises a first polypeptide chain and a second polypeptide chain, which method comprises:
- introducing into host cells:
- (i) a first population of vectors comprising nucleic acid encoding one or more of the first polypeptide chains which have been selected to have affinity for the predetermined target fused to a component of a secreted replicable genetic display package (RGDP) for display of the polypeptide chains at the surface of RGDPs;
- and
- (ii) a second population of vectors comprising nucleic acid encoding a genetically diverse population of the second polypeptide chain;
- the first vectors being packaged in infectious RGDPs and their introduction into host cells being by infection into host cells harboring the second vectors;
- or
- the second vectors being packaged in infectious RGDPs and their introducing into host cells being by infection into host cells harboring the first vectors;
- expressing the first and second polypeptide chains within the host cells to form a library of the SBP members displayed by RGDPs, at least one of the populations being expressed from nucleic acid that is capable of being packaged using the RGDP component, whereby the genetic materials of each the RGDP encodes a polypeptide chain of the SBP member displayed at its surface;
- and
- selecting members of the population for high-affinity binding to the predetermined target.
Dependencies are inferred from the claim text.