The application of an improved solid phase synthetic technique to the delineation of an antigenic site of apolipoprotein A-II

Pradip K. Bhatnagar, Simon J.T. Mao, Antonio Gotto, James T. Sparrow

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Previous studies have shown that the antigenic sites of human plasma high-density apolipoprotein A-II (apoA-II) are separate from their lipid-binding determinants in human high density lipoproteins (HDL). A specific radioimmunoassay has shown that three distinct antigenic sites are located in residues 4-23, 31-46, and 56-77; these studies suggested that an antigenic site might be restricted to residues 60-77 in the 56-77 fragment. To further delineate this site, we have developed a solid phase radioimmunoassay technique using an improved solid support on which selected sequences of peptides were synthesized, deprotected with HF, and the resulting peptidyl-resins tested for their capability of binding purified 125I-anti-apoA-II antibodies. Amino acid analyses and solid phase sequence analyses were performed to verify the sequence of the synthetic peptide on the solid support. Using this technique, 125I-anti-apoA-II antibodies had achieved 50% of maximal binding when residues 61-77 were attached to the solid support. The maximal binding was achieved by the addition of one more residue, Leu60, thus confirming our suggestion that a major antigenic site is located in residues 60-77. The binding to the peptidyl-resin was inhibited by a synthetic fragment corresponding to residues 60-77 indicating that the antibodies were specifically bound to the resin.

Original languageEnglish (US)
Pages (from-to)343-349
Number of pages7
JournalPeptides
Volume4
Issue number3
DOIs
StatePublished - 1983

Keywords

  • Antibodies
  • Antigenic sites
  • Apolipoprotein A-II
  • Improved resin
  • Peptide synthesis
  • Radioimmunoassay
  • Solid phase sequencing
  • Solid support
  • Synthetic peptides

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Physiology
  • Endocrinology
  • Biochemistry

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