CAJ | 학술논문

目的：运用辣根过氧化物酶标记JWA 单克隆抗体，并且将标记的抗体应用于ELISA法和western blot等免疫检测反应中。方法：运用高碘酸盐-四氢硼化钠氧化还原体系活化辣根过氧化物酶，活化的辣根过氧化物酶按1∶1标记JWA单克隆抗体；将HRP标记的抗体分别运用直接和双夹心ELISA测试；同时将标记单克隆抗体运用于western blot检测。结果：辣根过氧化物酶成功标记JWA单克隆抗体。其中HRP-JWA（4C9）抗体在直接法ELISA试验中和多肽的结合能力高于HRP-JWA（7C3）；其在450 nm处的最大吸光度值为0.96。双夹心ELISA实验中，预先包被JWA（4C9）单抗，检测抗体使用HRP-JWA（7C3）可以得到较好的实验结果，在450 nm处的最大吸光度值为1.30。在western blot实验HRP-JWA抗体浓度1.0μg·mL-1可以检测出SGC7901细胞中目的蛋白。结论：运用氧化还原方法成功标记JWA单克隆抗体，并可以初步应用于免疫反应的检测。
목적：운용랄근과양화물매표기JWA 단극륭항체，병차장표기적항체응용우ELISA법화western blot등면역검측반응중。방법：운용고전산염-사경붕화납양화환원체계활화랄근과양화물매，활화적랄근과양화물매안1∶1표기JWA단극륭항체；장HRP표기적항체분별운용직접화쌍협심ELISA측시；동시장표기단극륭항체운용우western blot검측。결과：랄근과양화물매성공표기JWA단극륭항체。기중HRP-JWA（4C9）항체재직접법ELISA시험중화다태적결합능력고우HRP-JWA（7C3）；기재450 nm처적최대흡광도치위0.96。쌍협심ELISA실험중，예선포피JWA（4C9）단항，검측항체사용HRP-JWA（7C3）가이득도교호적실험결과，재450 nm처적최대흡광도치위1.30。재western blot실험HRP-JWA항체농도1.0μg·mL-1가이검측출SGC7901세포중목적단백。결론：운용양화환원방법성공표기JWA단극륭항체，병가이초보응용우면역반응적검측。
Objective: To label JWA monoclonal antibody with horseradish peroxidase (HRP) and apply it in ELISA and western blot immune detection. Methods: Using sodium periodate and sodium borohydride redox reaction, HRP was activated; The activated HRP was used to mark JWA monoclonal antibody in tubes; The HRP labeled antibody was respectively applied in direct, sandwich ELISA and western blot de-tection. Results: HRP was successfully labeled on JWA monoclonal antibodies. In the direct method of ELISA test, HRP-JWA (4c9) antibodies combined polypeptide better than the combination of HRP-JWA (7c3); Its max OD450 was 0.96. While in double sandwich ELISA experiment, pre-coated by JWA (4c9) an-tibody, using the HRP-JWA (7c3) antibody as detected antibody could get good results, the max OD450 was 1.30. HRP-JWA could also identify the target protein in SGC7901 cells in western blot experiments. Conclusion: JWA antibody has been successfully marked with HRP, which could be used preliminarily in ELISA and western blot.