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J. Biochem, 1985, Vol. 98, No. 4 949-962
© 1985 Japanese Biochemical Society

other

Native and Non-Native Conformation-Specific Antibodies Directed to the Loop Region of Hen Egg-White Lysozyme

Hajime FUJIO1, Yutaka TAKAGAKI, Youn-Mun HA2, Elvira Missako DOI3, Amin SOEBANDRIO4 and Nobuo SAKATO

Department of Immunochemistry, Research Institute for Microbial Diseases, Osaka University Suita, Osaka 565

1 To whom requests for reprints should be addressed.

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.

2 Present address: Department of Microbiology, Medical School, Kyung Hee University, Hae Kee Dong, Dong Dae Mun Gu, Seoul, Korea.

3 Present address: Universidade Federal do Parana, Rua General Carneiro, 181 Curitiba Pr., Brazil.

4 Present address: Department of Microbiology, Medical Faculty University of Indonesia JL, Pegangsaan Timur 19, Jakarta Pusat, Indonesia.


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