Novales-Li P
Department of Pharmacology, University of Oxford, United Kingdom.
Biochem Biophys Res Commun. 1994 Sep 15;203(2):1076-82. doi: 10.1006/bbrc.1994.2292.
Using as antigen a 16 amino acid peptide corresponding to the C-terminus of soluble human brain acetylcholinesterase (AChE), four monoclonal antibodies were raised by the novel procedure of in vitro immunization. MAbs BMS-5, -6, -7, and -8 all demonstrated positive binding to the isoforms of cerebrospinal fluid (CSF) and foetal bovine serum (FBS) AChE. In contrast, no binding was seen with "membrane-bound" caudate nucleus and erythrocyte AChEs. With CSF samples from Alzheimer's patients, immunoblotting of isoelectric focussing (IEF) gels with the four mAbs failed to demonstrate any positive binding to the anomalous AChE band. Immunoblotting of SDS-reduced AChE showed binding of the four mAbs with "soluble" caudate nucleus and FBS AChE, but not with erythrocyte AChE. Hence, mAbs BMS-5, BMS-6, BMS-7 and BMS-8 all bind specifically to tetrameric AChE under non-denaturing and denaturing conditions.
使用对应于可溶性人脑乙酰胆碱酯酶（AChE）C末端的16个氨基酸肽作为抗原，通过体外免疫的新方法制备了四种单克隆抗体。单克隆抗体BMS-5、-6、-7和-8均显示与脑脊液（CSF）和胎牛血清（FBS）AChE的同工型呈阳性结合。相比之下，与“膜结合”的尾状核和红细胞AChE未见结合。用来自阿尔茨海默病患者的脑脊液样本，用这四种单克隆抗体对等电聚焦（IEF）凝胶进行免疫印迹，未能显示与异常AChE条带的任何阳性结合。对SDS还原的AChE进行免疫印迹显示，这四种单克隆抗体与“可溶性”尾状核和FBS AChE结合，但不与红细胞AChE结合。因此，单克隆抗体BMS-5、BMS-6、BMS-7和BMS-8在非变性和变性条件下均特异性结合四聚体AChE。
