Anti-epitope antibody, a novel site-directed antibody against human acetylcholinesterase  

作　　者：Xing-meiZHANG GangLIU Man-jiSUN 

机构地区：[1]InstituteofPharmacologyandToxicology,AcademyofMilitaryMedicalSciences,Beijing100850,China [2]CorrespondencetoProfMan-jiSUN

出　　处：《Acta Pharmacologica Sinica》2004年第4期431-435,共5页中国药理学报（英文版）

基　　金：Project supported by the "9th Five-Year" Military Medicine Health Science Foundation (No 96Z017).

摘　　要：AIM: To construct synthetic antigens using the epitope of human brainacetylcholinesterase (hbAChE) for induction and detection of the specific antibody against theepitope, and to analyse the immunogenicity of the antibody. METHODS: The epitope (RTVLVSMNYR, aminoacids 143-152) of hbAChE was chemically synthesized, coupled with the carrier protein keyhole limpethemocyanin (KLH) to construct an artificial immunogen (KLH-epitope), and injected into rabbits toraise antibody. The epitope conjugated with bovine serum albumin (BSA) was used as the detectionantigen. The specificity of the antibody was tested by enzyme-linked immunosorbent assay (ELISA) andWestern blotting. The immunoreaction between the anti-recombinant human butyrylcholinesterase(rhBChE) polyclonal antibody and the biotinylated-epitope was examined by indirect ELISA. RESULTS:The erythrocyte AChE, the hbAChE, rhBChE and the BSA-epitope all immunoreacted with the anti-epitopeantibody against the epitope (143-152) of hbAChE, whereas the torpedo AChE did not. CONCLUSION: ThehbAChE, the human erythrocyte AChE and hBChE share the conservative antigenic epitope RTVLVSMNYR,hence they can all immunoreact with the anti-epitope antibody. Since the epitope of hbAChE is lesssimilar with the aligned amino acid sequences of AChE of Torpedo californica or Torpedo marmorata,there is not any immunoreactivity between them. The R, M, and N residues in the epitope seem to benecessary radicals for the conservation of antigenicity.AIM: To construct synthetic antigens using the epitope of human brainacetylcholinesterase (hbAChE) for induction and detection of the specific antibody against theepitope, and to analyse the immunogenicity of the antibody. METHODS: The epitope (RTVLVSMNYR, aminoacids 143-152) of hbAChE was chemically synthesized, coupled with the carrier protein keyhole limpethemocyanin (KLH) to construct an artificial immunogen (KLH-epitope), and injected into rabbits toraise antibody. The epitope conjugated with bovine serum albumin (BSA) was used as the detectionantigen. The specificity of the antibody was tested by enzyme-linked immunosorbent assay (ELISA) andWestern blotting. The immunoreaction between the anti-recombinant human butyrylcholinesterase(rhBChE) polyclonal antibody and the biotinylated-epitope was examined by indirect ELISA. RESULTS:The erythrocyte AChE, the hbAChE, rhBChE and the BSA-epitope all immunoreacted with the anti-epitopeantibody against the epitope (143-152) of hbAChE, whereas the torpedo AChE did not. CONCLUSION: ThehbAChE, the human erythrocyte AChE and hBChE share the conservative antigenic epitope RTVLVSMNYR,hence they can all immunoreact with the anti-epitope antibody. Since the epitope of hbAChE is lesssimilar with the aligned amino acid sequences of AChE of Torpedo californica or Torpedo marmorata,there is not any immunoreactivity between them. The R, M, and N residues in the epitope seem to benecessary radicals for the conservation of antigenicity.

关 键 词：ACETYLCHOLINESTERASE BUTYRYLCHOLINESTERASE EPITOPES antibodies enzyme-linked immunosorbent assay Western blotting 

分 类 号：Q55[生物学—生物化学]