| Cysticercosis cellulosae is one of the most important Znonotic Parasitic diseases in our country. Itcauses great economic losses, and has serious effect on public health. Human is not only a intermediatehost but also a terminal host. Cysticercus is a kind of parasite which often lives in manyorgans such asbrain, eyes, heart and so on. When living innervous system,it can cause serious disease,for examplecerebral cysticercosis which does great harm to health of human. The aim of our present study was toprepare thehybridoma cell lines which can stably produce McAb against Cysticercus Cellulosae Scolexantigen and the McAbs, to identify the cell lines' biological characteristics. Double-antibody sandwichELISA method for detecting relative quantity of Cysticercus Cellulosae Scolex protein were establishedand applicated.Methods: SephadexG-200 chromatographic technology was performed for purifying cysticercosisScolex rude antigen (CS), The BALB/c mice were immunized by cysticercosis Scolex purified antigenfor 4 times. The immune interval was two weeks. And the mouse was attacked by Scolex purifiedantigen three days before the hybridization. The spleen cells of the immunized mice were hybridizedwith myeloma cell line Sp2/0 cells by PEG. The hybridized cells were selected in HAT. The positivehybridoma cells secreting the antibody were determined by the indirect ELISA. Through cloningculture, stable hybridoma cells lines secreting anti-Scolex McAbs were obtained.Anti-Scolex McAbpositive hybridoma cell line cells were injected into the abdominal cavity of BALB/c mice. Titers ofMcAbs were tested by the indirect ELISA. Saturated ammonium sulfate (SAS) were obtainthepreliminarily purified McAb (SAS-McAb). The purity of the purified McAb was identified bysodium dodecylsulphate-polycrylamide gel electrophorisis (SDS—PAGE). Titers of SAS-McAbs weretested by the indirect ELISA. We tested specific of the mAbs by the indirect ELISA. The New Zealandrabbits were immunized with the cysticercosis Scolex purified antigen to prepare the serum polyclonalantibody (PcAb). The PcAb of IgG was purified by the SAS precipitation from serum PcAb. TheSandwich ELISA Was established with purified single McAb-PcAb was used to detect thecysticercosis Scolex antigen at respectively, which different dilution concentration to determine thesensitivity. The system was used to check the cysticercosis cyst fluid and cysticercosis capsule wallto specificity for cysticercosis Scolex antigen. Result: Three hybridoma cell lines secreting McAb against cysticercosis Scolex antigen wereprimary obtained by cell hybridation, HAT selection, cloning culture and antibody screening which werenamed as 2C6,3A5和4G10, respectively. The McAb can be secreted stably by these cell lines after thirtysubcultures in vitro and thawed after frozen for 3 months. The titers of ascites from the mice injectedwith 2C6 was 1: 6.4×10~4, 3A5 and 4G10 were 1: 1.6×10~3,1: 3.2×10~3 respectively. Within three cell lines,the 2C6 secrets IgG1, and the others secret IgG2a. Cysticercosis Scolex ELISA is an enzymaticallyamplified "two-step" sandwich-type immunoassay. In the assay, Standards,controls and unknow serumsamples are incubated in microtitration wells which have been coated with anti-Cysticercosis Scolexcapture antibody(2C6) incubated. After incubation and washing the wells are treated with another anti-Cysticercosis Scolex detection(anti-serum) antibody. Anti-rabbit of antibody labeled with the enzymehorseradish peroxidase(HRP) was the second antibody, After a second incubation and washing step,thewells are incubated with the substrate OPD.An acidic stopping solution is then added and the degrer ofenzymatic turnover of the sunstrate is determined by single wavelength absorbance measurement at 490nm.The concentration logarithm takes on linear correlation with OD490nm at the range of CysticercosisScolex content from 1ng/mL～10μg/mL.The content of unkown sample is computed comparied withstandards.This assay method is simple and sensitive.
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