Fujio H, Takagaki Y, Ha Y M, Doi E M, Soebandrio A, Sakato N
J Biochem. 1985 Oct;98(4):949-62. doi: 10.1093/oxfordjournals.jbchem.a135375.
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.
在传统的豚鼠抗鸡卵清溶菌酶（HEL）抗血清中区分出了两种类型的抗体。两种抗体的特异性均针对环I区域（主要针对Cys64 - Cys80环），但它们与天然HEL的反应性不同。一种抗体与HEL及HEL的环肽反应，但不与环肽的完全还原和羧甲基化形式反应（天然构象特异性抗体：NC - Ab）。另一方面，另一种抗体不与HEL反应，但与环肽以及环肽的完全还原和羧甲基化形式反应（非天然构象特异性抗体：NNC - Ab）。通过两种不同免疫方法获得的豚鼠抗HEL混合抗血清中环I反应性抗体组分中NNC - Ab的百分比约为25%。由于NNC - Ab对环相关肽的亲和力比NC - Ab对相同肽的亲和力高一个数量级，因此在评估天然蛋白质中的抗原决定簇时需要谨慎。用Ploop I. II [序列57 - 107（Cys64 - Cys80，Cys76 - Cys94）]免疫豚鼠可诱发一群抗体，其特异性与传统抗HEL抗血清中NNC - Ab型抗环I抗体的特异性相似但不完全相同。
