Characterization of four distinct monoclonal antibodies specific to BmK AS-1, a novel scorpion bioactive polypeptide.
Four monoclonal antibodies designed as 2#, 3#, 4# and 5# have been raised against a novel bioactive polypeptide BmK AS-1 purified from the Chinese scorpion Buthus martensi Karsch. All of these antibodies exhibited specific affinity with antigen by ELISA and Biosensor assay. Western blot analysis showed that 3# and 4# were able to recognize the denatured antigen, but not 2# and 5#. These antibodies could cross-react with BmK AS, but not with other types of BmK neurotoxins such as BmK I (an alpha-like toxin) and BmK IT (an excitatory insect-selective toxin), and in which only 5# can partially react with BmK IT2 (a depressant insect-selective toxin). Immunocytochemical staining demonstrated that 3#, 4# and 5# antibodies can visualize the antigen bound to the membrane of SK-N-SH neuroblast cells, with the exception of 2#. This suggests that either conformation alteration of receptor binding might be prone to nonvisualization or the epitope recognized by antibody 2# might be overlapped with receptor binding sites of antigen. The antibodies developed in the study should provide powerful new tools for investigating the structure/function relationship and pharmacological mechanism of scorpion neurotoxins.
Jia
L Y
LY
Shanghai Institute of Physiology, Chinese Academy of Sciences, Shanghai Research Center of Life Sciences, People's Republic of China. yhji@server.shcnc.ac.cn
Xie
H F
HF
Ji
Y H
YH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D015374:Biosensing Techniques / Q000379:methods; D015153:Blotting, Western; D003429:Cross Reactions / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009447:Neuroblastoma / Q000276:immunology / Q000473:pathology; D009498:Neurotoxins / Q000276:immunology; D010455:Peptides / Q000276:immunology* / Q000302:isolation & purification; D012604:Scorpion Venoms / Q000276:immunology* / Q000302:isolation & purification; D012605:Scorpions / Q000276:immunology*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; C119408:KAS-1 toxin, Buthus martensi; D009498:Neurotoxins; D010455:Peptides; D012604:Scorpion Venoms