A synthetic strategy for simultaneous purification-conjugation of antigenic peptides

Anal Biochem. 1989 Sep;181(2):389-95. doi: 10.1016/0003-2697(89)90260-1.

Abstract

A novel approach to the preparation of immunopeptide-carrier protein conjugates of improved chemical definition based on a solid-phase synthetic protocol combining incorporation of a Cys(Npys) N-terminal residue with systematic acetylation after every coupling, is described. The potential of the method is demonstrated in a synthesis of the tridecapeptide (Npys)-Cys-Val-Asn-Tyr-Ile-Arg-Lys-Arg-Ser-Leu-Gln-Thr-Val-OH in which the main product is purposely contaminated by a number of shorter truncated sequences resulting from intentionally defective couplings. From this peptide crude, and rather independently of its complexity, the target sequence can be selectively recovered and attached to a carrier molecule through a disulfide bond formed by reaction of the Npys-protected cysteine residue and a thiol function in the carrier. The process can be properly named purification-by-conjugation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Antigens / analysis*
  • Borohydrides
  • Carrier Proteins / analysis
  • Carrier Proteins / chemical synthesis
  • Chromatography, High Pressure Liquid
  • Hemocyanins
  • Molecular Sequence Data
  • Peptide Fragments / analysis
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / isolation & purification
  • Peptides / chemical synthesis*
  • Peptides / immunology
  • Peptides / isolation & purification
  • Protein Binding
  • Sepharose
  • Serum Albumin, Bovine / analysis

Substances

  • Antigens
  • Borohydrides
  • Carrier Proteins
  • Peptide Fragments
  • Peptides
  • Serum Albumin, Bovine
  • sodium borohydride
  • Sepharose
  • Hemocyanins