CAJ | 학술논문

为了研制氯霉素单克隆抗体,建立氯霉素的简便有效的检测方法.采用碳二亚胺(EDC)方法制备氯霉素免疫抗原和包被抗原,经SDS-PAGE凝胶电泳,三硝基苯磺酸(TNBS)法鉴定表明偶联成功.采用制备的抗原CAP-HS-BSA((CAP-HS,氯霉素琥珀酸酯;BSA,牛血清白蛋白)免疫Balb/c小鼠,通过杂交瘤技术获得了2株抗氯霉素的杂交瘤细胞株,细胞株体外传代和冻存复苏后抗体分泌稳定.经体内诱生法产生腹水,ELISA检测纯化腹水的效价为1:10~6.抗体的亲和常数分别为:1.13×10~(10) L/mol,1.41×10~(10) L/mol.ELISA检测显示这2株单克隆抗体与氯霉素琥珀酸交叉反应率为300%,与其他抗生素及结构类似物的交叉反应小,表明该单抗可满足建立免疫学检测方法的需要和开发应用的要求.
위료연제록매소단극륭항체,건립록매소적간편유효적검측방법.채용탄이아알(EDC)방법제비록매소면역항원화포피항원,경SDS-PAGE응효전영,삼초기분광산(TNBS)법감정표명우련성공.채용제비적항원CAP-HS-BSA((CAP-HS,록매소호박산지;BSA,우혈청백단백)면역Balb/c소서,통과잡교류기술획득료2주항록매소적잡교류세포주,세포주체외전대화동존복소후항체분비은정.경체내유생법산생복수,ELISA검측순화복수적효개위1:10~6.항체적친화상수분별위:1.13×10~(10) L/mol,1.41×10~(10) L/mol.ELISA검측현시저2주단극륭항체여록매소호박산교차반응솔위300%,여기타항생소급결구유사물적교차반응소,표명해단항가만족건립면역학검측방법적수요화개발응용적요구.
In order to prepare momclonal antibody(Mab) against chlormphenicol(CAP) for CAP detection, CAP was conjugated with bovine serum albumin(BSA) and ovalbumin(OVA) to prepare immune antigen and detection antigen by the 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC) method respectively, then the artificial antigens were detected by SDS-PAGE, and the residual amount of ε-amino groups was determined by the TNBS method. The result showed it was successfully synthesized. Balb/c mice were immunized with conjugated CAP-HS-BSA, and two strains of hybridomas which secret high titer antibodies against CAP were cloned and then ascitic fluids were made, the titers of ascites were 1×10~6 by ELISA and the affinity constant was 1.13×10~(10) L/mol and 1.41×10~(10) L/mol, respectively. The number of chromosome of the hybridoma cell lines was 90～104. The cross reaction rate of Mab against CAP and CAP-HS was 300%, and the Mab did not have cross reactivity with other antibiotics. The result showed the Mab can be used for the detection of CAP.