Exploring antibody polyspecificity using synthetic combinatorial libraries.
Extensive mapping studies for seven antigen-antibody interactions have been carried out using both individual analogs and peptide libraries. With competitive ELISA, these studies have revealed that monoclonal antibodies exhibit a broad range of specificities, from antibodies that recognize only conservative substitutions for 1-2 positions of the antigenic determinant, to antibodies that recognize sequences that are completely unrelated to the parent antigen with comparable affinities. Synthetic combinatorial libraries, containing millions of peptide sequences, permit a more systematic and rapid evaluation of the extent of multiple-binding specificities of monoclonal antibodies than individual analogs. The peptide libraries used here comprise mixtures of compounds having specifically defined positions and mixture positions. The same diversity of sequences in different formats, which differ by the numbers of positions singularly defined and different locations defined within the sequence, can be examined. Comparison of the screening results, selection criteria of the most active mixtures, and different approaches used for the deconvolution of active individual compounds are discussed. Synthetic combinatorial libraries greatly facilitate the understanding of antigen-antibody interactions at the amino acid level and will assist in the development of improved immunodiagnostics.
Appel
J R
JR
Torrey Pines Institute for Molecular Studies, San Diego, CA 92121, USA.
Buencamino
J
J
Houghten
R A
RA
Pinilla
C
C
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry; D000918:Antibody Specificity; D000939:Epitopes / Q000276:immunology; D005340:Fibrinogen / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D008969:Molecular Sequence Data; D009842:Oligopeptides; D018739:Oncogene Proteins v-erbB / Q000276:immunology; D019151:Peptide Library; D010455:Peptides / Q000276:immunology; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins / Q000276:immunology; D051578:Proto-Oncogene Proteins c-fes; D051153:Wnt Proteins; D029961:Zebrafish Proteins
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006514:Hepatitis B Surface Antigens; D009842:Oligopeptides; D018739:Oncogene Proteins v-erbB; D019151:Peptide Library; D010455:Peptides; D011518:Proto-Oncogene Proteins; D051153:Wnt Proteins; D029961:Zebrafish Proteins; D005340:Fibrinogen; C085706:FLAG peptide; D011505:Protein-Tyrosine Kinases; D051578:Proto-Oncogene Proteins c-fes