Native and non-native conformation-specific antibodies directed to the loop region of hen egg-white lysozyme.
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 carboxymethylated 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 carboxymethylated 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.
Fujio
H
H
Takagaki
Y
Y
Ha
Y M
YM
Doi
E M
EM
Soebandrio
A
A
Sakato
N
N
D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D002645:Chickens; D004220:Disulfides; D000939:Epitopes; D005260:Female; D006168:Guinea Pigs; D009113:Muramidase / Q000276:immunology*; D011487:Protein Conformation
D004220:Disulfides; D000939:Epitopes; D009113:Muramidase