Searching for peptide ligands with an epitope library.

Tens of millions of short peptides can be easily surveyed for tight binding to an antibody, receptor or other binding protein using an "epitope library." The library is a vast mixture of filamentous phage clones, each displaying one peptide sequence on the virion surface. The survey is accomplished by using the binding protein to affinity-purify phage that display tight-binding peptides and propagating the purified phage in Escherichia coli. The amino acid sequences of the peptides displayed on the phage are then determined by sequencing the corresponding coding region in the viral DNA's. Potential applications of the epitope library include investigation of the specificity of antibodies and discovery of mimetic drug candidates.

[1]  L. Day Conformations of single-stranded DNA and coat protein in fd bacteriophage as revealed by ultraviolet absorption spectroscopy. , 1969, Journal of molecular biology.

[2]  T. McCutchan,et al.  Immunogenicity and epitope mapping of foreign sequences via genetically engineered filamentous phage. , 1988, The Journal of biological chemistry.

[3]  W. J. Dower,et al.  High efficiency transformation of E. coli by high voltage electroporation , 1988, Nucleic Acids Res..

[4]  D W TALMAGE,et al.  Immunological Specificity , 1959, Science.

[5]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[6]  G. P. Smith,et al.  Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. , 1988, Gene.

[7]  K. Mullis,et al.  Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. , 1987, Methods in enzymology.

[8]  G. P. Smith,et al.  Filamentous phage assembly: morphogenetically defective mutants that do not kill the host. , 1988, Virology.

[9]  Influence of protein flexibility and peptide conformation on reactivity of monoclonal anti-peptide antibodies with a protein alpha-helix. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[10]  W. Konigsberg,et al.  On the specificity of antibodies. , 1975, Science.

[11]  R. Lerner,et al.  Preliminary crystallographic data and primary sequence for anti-peptide Fab' B13I2 and its complex with the C-helix peptide from myohemerythrin. , 1989, The Journal of biological chemistry.

[12]  Thomas A. Kunkel,et al.  Rapid and efficient site-specific mutagenesis without phenotypic selection. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[13]  J. Devlin,et al.  Random peptide libraries: a source of specific protein binding molecules. , 1990, Science.

[14]  W. Wickner Asymmetric orientation of a phage coat protein in cytoplasmic membrane of Escherichia coli. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[15]  R. Porter,et al.  Synthetic peptides as antigens. , 1986, Ciba Foundation symposium.