Sequence and structure specific antibodies from phage display libraries

Abstract

A large combinatorial phage display library was panned against five nucleic acid antigens, calf thymus DNA, poly[d(GC)], poly[d(AT)], poly(dA) · poly(dT) and poly(rA) · poly(dT). After the third and fourth rounds of panning, many positive clones were selected against poly[d(GC)], poly(dA) · poly(dT) and poly(rA) · poly(dT). The specificity of these antibodies was tested by both direct and competitive solid phase radioimmune assays. All the clones derived from panning with poly[d(GC)] were non-specific and bound to all nucleic acids. The poly(rA) · poly(dT) derived clones were specific for single-stranded nucleic acids, with some sequence preferences, and the poly(dA) · poly(dT) derived clones showed considerable specificity for this antigen. The sequences of these phage-derived antibodies showed no similarities with DNA-binding antibodies from other sources. Even after six rounds of panning no positive clones were detected which bound to poly[d(AT)] and after seven rounds only two were derived from panning with calf thymus DNA. Therefore, sequence-and structure specific antibodies can be recovered from phage display libraries but not all sequences may be represented in the repertoire.