论文部分内容阅读
用以检测抗原特异表位的竞争结合抗体法(CABA),如用粗制蛋白抗原做时,对某一特异性抗原表位的单克隆抗体的反应可被受检血清中的抗体竞争所抑制。这一假阳性反应可能是受检血清中多克隆抗体对粗制抗原的多表位发生反应,使邻近的这一特异性表位与单克隆抗体的结合受阻,而并非血清中确有这一特异抗体。如改用提纯的天然蛋白或重组蛋白抗原呈阴性反应就能证实这种假阳性。作者于感染麻风菌前和感染后3个月一次采集犰狳静脉血,还有四位麻风患者的血,用麻风菌的65KD蛋白做免疫印迹和CABA检查。CABA是用100μl麻风菌胞壁成份(含65KD蛋白的粗制抗原)和重组的65KD蛋白抗原包被多孔微滴板,经洗涤封闭后,加入50μl放射碘标记的IIIE_9单克隆抗体(麻
The competitive binding antibody method (CABA) to detect antigen-specific epitopes, as done with a crude protein antigen, can be counteracted by the competition of antibodies in the sera tested for the response to a monoclonal antibody to a particular epitope . This false-positive reaction may be due to polyclonal antibodies in the sera tested against the polyepitope of the crude antigen, blocking the binding of this specific epitope to the monoclonal antibody, rather than in the serum Specific antibody. Such as the use of purified native protein or recombinant protein antigen negative reaction can confirm this false positive. The authors collected armadillo venous blood before infecting leprosy and 3 months after infection, as well as blood from four leprosy patients, immunoblotting and CABA using a 65 kD protein of leprosy. CABA was coated with 100 microliters of lepromatous cell wall composition (crude antigen containing 65KD protein) and recombinant 65KD protein antigen coated porous microtiter plate, washed and blocked, then added 50μl of radioactive iodine-labeled monoclonal antibody IIIE_9