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Antigenic and Allergenic Determinants of Ovalbumin
Author(s) -
ELSAYED S.,
HOLEN E.,
HAUGSTAD M. B.
Publication year - 1988
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1988.tb02386.x
Subject(s) - ovalbumin , chemistry , peptide , edman degradation , epitope , amino acid , high performance liquid chromatography , chromatography , size exclusion chromatography , peptide synthesis , peptide sequence , microscale thermophoresis , antigen , biochemistry , biology , immunology , gene , enzyme
The decapeptide 1‐10 of hen's egg ovalbumin (OA), deduced from the known amino acid sequence (Gly‐Ser‐Ile‐Gly‐Ala‐Ala‐Ser‐Met‐Glu‐Phe), was synthesized by Merrifield solid phase peptide synthesis with a yield of >70%. The completeness of the insertion of amino acids during synthesis was monitored by the amino acid compositions of the peptide‐resin, prior to coupling of the preceding residue. The linearity of the synthesis was supported by dansyl Edman degradation and detection of three NH 2 terminal residues 5‐dimethylamino‐naphthdine‐1‐sulphonyl (DNS)‐Gly, ‐Ser, ‐Ile, respectively. The peptide was purified by gel filtration chromatography and analytical reversed‐phase high‐performance liquid chromatography (HPLC). The homogeneity of the preparation was calculated both from the amino acid analysis and by integrating the peaks of HPLC to be >83%. The antigenicity of the purified (P2) peptide could be detected by precipitation inhibition with the nephelometric technique. The decapeptide could also specifically react with functional structures on reaginic IgE molecule from the sera of individuals allergic to eggs, inhibiting its further binding to ovalbumin An in‐vivo experiment using direct skin test on two patients allergic to eggs showed no activity, rendering further testing unnecessary. The results suggest that the decapeptide of the NH 2 terminal segment of OA encompasses an Ig‐binding haptenic epitope.