Sequence and structure specific antibodies from phage display libraries.
A large combinatorial phage display library was panned against five nucleic acid antigens, calf thymus DNA, poly[d(GC)], poly[d(AT)], poly(dA) x poly(dT) and poly(rA) x poly(dT). After the third and fourth rounds of panning, many positive clones were selected against poly[d(GC)], poly(dA) x poly(dT) and poly(rA) x 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) x poly(dT) derived clones were specific for single-stranded nucleic acids, with some sequence preferences, and the poly(dA) x 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.
Tanha
J
J
Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.
Forsyth
G
G
Schorr
P
P
Crosby
W
W
Lee
J S
JS
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D001435:Bacteriophages; D004247:DNA / Q000276:immunology*; D004268:DNA-Binding Proteins / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D019151:Peptide Library; D011061:Poly A / Q000276:immunology; D011071:Poly T / Q000276:immunology; D011067:Poly dA-dT / Q000276:immunology; D011089:Polydeoxyribonucleotides / Q000276:immunology; D011863:Radioimmunoassay; D013329:Structure-Activity Relationship
D000906:Antibodies; D004268:DNA-Binding Proteins; D019151:Peptide Library; D011089:Polydeoxyribonucleotides; D011061:Poly A; D011071:Poly T; D011067:Poly dA-dT; C008950:poly A-T; C019530:poly(dC-dG); C084838:polydeoxyadenylic acid-polythymidylic acid; D004247:DNA