Native and non-native conformation-specific antibodies directed to the loop region of hen egg-white lysozyme

Hajime Fujio, Yutaka Takagaki, Youn Mun Ha, Elvira Missako Doi, Amin Subandrio W. Kusumo, Nobuo Sakato

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Two types of antibodies were differentiated in conventional Guinea pig anti-hen egg-white lysozyme (HEL) antisera. The specificities of both antibodies were directed to the loop I region (mainly directed to Cys64-Cys80 loop) but the antibodies were distinct in respect of reactivities with native HEL. One type of antibody reacted with HEL and loop-peptides of HEL but not with the completely reduced and car-boxymethylated form of loop-peptides (native conformation specific antibody: NC-Ab). On the other hand, the other type of antibody did not react with HEL but reacted with loop-peptides and also with the completely reduced and carboxymeth-ylated form of loop-peptides (non-native conformation specific antibody: NNC-Ab). The percentage of NNC-Ab in loop I reactive antibody fraction from pooled Guinea pig anti-HEL antisera obtained by two different immunization methods was about 25%. Since the affinities of the NNC-Ab to loop-related peptides were higher by one order of magnitude than those of the NC-Ab to the same peptides, care is necessary in evaluating antigenic determinants in native protein.The immunization of Guinea pigs with Ploop I II [sequence 57-107 (Cys64-Cys80, Cys76-Cys94)] evoked an antibody population having specificity similar to but not identical with that of the NNC-Ab type anti-loop I antibody in conventional anti-HEL antisera.

Original languageEnglish
Pages (from-to)949-962
Number of pages14
JournalJournal of Biochemistry
Volume98
Issue number4
DOIs
Publication statusPublished - 1 Jan 1985

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