CAJ | 학술논문

用人工合成的氯霉素-卵清蛋白(CAP-OVA)免疫BALB/c小鼠,通过杂交瘤技术筛选出1株能稳定传代并分泌抗氯霉素(CAP)单克隆抗体(McAb)的杂交瘤细胞4C9,并以此制备腹水单抗.经间接竞争ELISA(ciELISA)检测,该单抗亚类重链类型是IgG1,轻链为κ类型,单抗腹水效价为1:1×10~6,与甲砜霉素、氟甲砜霉素以及其他常见抗生素的交叉反应率小于0.01%.用过碘酸钠氧化法合成CAP酶标记单抗,建立了CAP直接竞争ELISA(cdELISA)方法.此法检测的线性范围为0.1～100 ng·mL~(-1),半数抑制浓度(IC_(50))为5.81 ng·L~(-1).添加回收实验表明所建立的CAP cdELISA方法检测限达到0.1 ng·L~(-1),对比试验表明其检测灵敏度与商品试剂CAP ELISA基本相当.
용인공합성적록매소-란청단백(CAP-OVA)면역BALB/c소서,통과잡교류기술사선출1주능은정전대병분비항록매소(CAP)단극륭항체(McAb)적잡교류세포4C9,병이차제비복수단항.경간접경쟁ELISA(ciELISA)검측,해단항아류중련류형시IgG1,경련위κ류형,단항복수효개위1:1×10~6,여갑풍매소、불갑풍매소이급기타상견항생소적교차반응솔소우0.01%.용과전산납양화법합성CAP매표기단항,건립료CAP직접경쟁ELISA(cdELISA)방법.차법검측적선성범위위0.1～100 ng·mL~(-1),반수억제농도(IC_(50))위5.81 ng·L~(-1).첨가회수실험표명소건립적CAP cdELISA방법검측한체도0.1 ng·L~(-1),대비시험표명기검측령민도여상품시제CAP ELISA기본상당.
Through immunization of BALB/c mice with synthesized CAP-ovalbumin (CAP-OVA), a hybridoma cell line, the 4C9 which could produce monoclonal antibody against chloramphenicol (CAP) and propagate stably, was established. With the myeloma cells, the ascites containing monclonal antibody (McAb) were prepared. The indirect competitive ELISA (ciELISA) test revealed the heavy chain and light chain of McAb were IgG1 and κ, respectively. The ELISA titer of ascites was 1:1×10~6. The percentages of cross-activity to thiamphenicol, florfenicol and other antibiotics were all less than 0.01%. A horseradish peroxidase (HRP)-labeled antibody was synthesized by the sodium periodate reaction, then a direct competitive enzyme-linked immunosorbent assay (cdELISA) was developed. Quantization of the CAP was linear from 0.1 to 100 ng·mL~(-1), and the hemi-inhibitory concentration (IC_(50)) was 5.81 ng·L~(-1). The recovery test showed that the detection limit for CAP was 0.1 ng·L~(-1) in cdELISA. Comparative test showed that detected sensibility of the monoclonal antibody was nearly equivalent to the commercial CAP ELISA kit.