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Proc Natl Acad Sci U S A. Aug 1985; 82(15): 5131–5135.
PMCID: PMC390513

General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids.

Abstract

A novel yet simple method is described that facilitates the synthesis of large numbers of peptides to the extent that the synthesis process need no longer be the limiting factor in many studies involving peptides. By using the methods described, 10-20 mg of 248 different 13-residue peptides representing single amino acid variants of a segment of the hemagglutinin protein (HA1) have been prepared and characterized in less than 4 weeks. Through examination of the binding of these analogs to monoclonal antibodies raised against residues 75-110 of HA1, it was found that a single amino acid, aspartic acid at position 101, is of unique importance to the interaction. Two other residues, aspartic acid-104 and alanine-106, were found to play a lesser but significant role in the binding interaction. Other single positional residue variations appear to be of little or no importance.

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Selected References

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  • Lerner RA. Tapping the immunological repertoire to produce antibodies of predetermined specificity. Nature. 1982 Oct 14;299(5884):593–596. [PubMed]
  • Sutcliffe JG, Shinnick TM, Green N, Liu FT, Niman HL, Lerner RA. Chemical synthesis of a polypeptide predicted from nucleotide sequence allows detection of a new retroviral gene product. Nature. 1980 Oct 30;287(5785):801–805. [PubMed]
  • Green N, Alexander H, Olson A, Alexander S, Shinnick TM, Sutcliffe JG, Lerner RA. Immunogenic structure of the influenza virus hemagglutinin. Cell. 1982 Mar;28(3):477–487. [PubMed]
  • Bittle JL, Houghten RA, Alexander H, Shinnick TM, Sutcliffe JG, Lerner RA, Rowlands DJ, Brown F. Protection against foot-and-mouth disease by immunization with a chemically synthesized peptide predicted from the viral nucleotide sequence. Nature. 1982 Jul 1;298(5869):30–33. [PubMed]
  • East IJ, Todd PE, Leach SJ. On topographic antigenic determinants in myoglobins. Mol Immunol. 1980 Apr;17(4):519–525. [PubMed]
  • Wilson IA, Niman HL, Houghten RA, Cherenson AR, Connolly ML, Lerner RA. The structure of an antigenic determinant in a protein. Cell. 1984 Jul;37(3):767–778. [PubMed]
  • McMillan S, Seiden MV, Houghten RA, Clevinger B, Davie JM, Lerner RA. Synthetic idiotypes: the third hypervariable region of murine anti-dextran antibodies. Cell. 1983 Dec;35(3 Pt 2):859–863. [PubMed]
  • Geysen HM, Meloen RH, Barteling SJ. Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc Natl Acad Sci U S A. 1984 Jul;81(13):3998–4002. [PMC free article] [PubMed]
  • Geysen HM, Barteling SJ, Meloen RH. Small peptides induce antibodies with a sequence and structural requirement for binding antigen comparable to antibodies raised against the native protein. Proc Natl Acad Sci U S A. 1985 Jan;82(1):178–182. [PMC free article] [PubMed]
  • Niman HL, Houghten RA, Walker LE, Reisfeld RA, Wilson IA, Hogle JM, Lerner RA. Generation of protein-reactive antibodies by short peptides is an event of high frequency: implications for the structural basis of immune recognition. Proc Natl Acad Sci U S A. 1983 Aug;80(16):4949–4953. [PMC free article] [PubMed]
  • Veber DF, Holly FW, Paleveda WJ, Nutt RF, Bergstrand SJ, Torchiana M, Glitzer MS, Saperstein R, Hirschmann R. Conformationally restricted bicyclic analogs of somatostatin. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2636–2640. [PMC free article] [PubMed]
  • Chen PP, Houghten RA, Fong S, Rhodes GH, Gilbertson TA, Vaughan JH, Lerner RA, Carson DA. Anti-hypervariable region antibody induced by a defined peptide: an approach for studying the structural correlates of idiotypes. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1784–1788. [PMC free article] [PubMed]
  • Klipstein FA, Engert RF, Houghten RA, Rowe B. Enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxin. J Clin Microbiol. 1984 Jun;19(6):798–803. [PMC free article] [PubMed]
  • Chen PP, Fong S, Houghten RA, Carson DA. Characterization of an epibody. An antiidiotype that reacts with both the idiotype of rheumatoid factors (RF) and the antigen recognized by RF. J Exp Med. 1985 Feb 1;161(2):323–331. [PMC free article] [PubMed]
  • Sutcliffe JG, Milner RJ, Shinnick TM, Bloom FE. Identifying the protein products of brain-specific genes with antibodies to chemically synthesized peptides. Cell. 1983 Jul;33(3):671–682. [PubMed]
  • Freidinger RM, Veber DF, Perlow DS, Brooks JR, Saperstein R. Bioactive conformation of luteinizing hormone-releasing hormone: evidence from a conformationally constrained analog. Science. 1980 Nov 7;210(4470):656–658. [PubMed]
  • Atherton E, Clive DL, Sheppard RC. Letter: Polyamide supports for polypeptide synthesis. J Am Chem Soc. 1975 Oct 29;97(22):6584–6585. [PubMed]
  • Houghten RA, Chang WC, Li CH. Human beta-endorphin: synthesis and characterization of analogs iodinated and tritiated at tyrosine residues 1 and 27. Int J Pept Protein Res. 1980 Oct;16(4):311–320. [PubMed]
  • Houghten RA, Ostresh JM, Klipstein FA. Chemical synthesis of an octadecapeptide with the biological and immunological properties of human heat-stable Escherichia coli enterotoxin. Eur J Biochem. 1984 Nov 15;145(1):157–162. [PubMed]
  • Wieland T, Birr C, Flor F. Symmetrical Boc-amino acid anhydrides for economical peptide syntheses on a solid phase. Angew Chem Int Ed Engl. 1971 May;10(5):336–336. [PubMed]
  • Gorman JJ. An apparatus for simultaneous manual solid-phase synthesis of multiple peptide analogs. Anal Biochem. 1984 Feb;136(2):397–406. [PubMed]
  • Jou WM, Verhoeyen M, Devos R, Saman E, Fang R, Huylebroeck D, Fiers W, Threlfall G, Barber C, Carey N, et al. Complete structure of the hemagglutinin gene from the human influenza A/Victoria/3/75 (H3N2) strain as determined from cloned DNA. Cell. 1980 Mar;19(3):683–696. [PubMed]

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