Isolation of anti-angiogenesis antibodies from a large combinatorial repertoire by colony filter screening.

ABSTRACT: We describe here a method, based on iterative colony filter screening, for the rapid isolation of binding specificities from a large synthetic repertoire of human antibody fragments in single-chain Fv configuration. Escherichia coli cells, expressing the library of antibody fragments, are grown on a porous master filter, in contact with a second filter coated with the antigen, onto which antibodies secreted by the bacteria are able to diffuse. Detection of antigen binding on the second filter allows the recovery of a number of E.coli cells, including those expressing the binding specificity of interest, which can be submitted to a second round of screening for the isolation of specific monoclonal antibodies. We tested the methodology using as antigen the ED-B domain of fibronectin, a marke

SUBMITTER: Giovannoni L 

PROVIDER: S-EPMC29740 | biostudies-literature | 2001 Mar

REPOSITORIES: biostudies-literature