Furlong R A, Welch A N, Peake I R
Br J Haematol. 1987 Jul;66(3):341-8. doi: 10.1111/j.1365-2141.1987.tb06921.x.
Two monoclonal antibodies (McAb) termed 12A4 and 19C1 have been raised against human factor VIII. In immunoassays 12A4 bound to factor VIII antigen (VIII:Ag) in plasma but not serum whilst 19C1 bound to VIII:Ag in both plasma and serum. Both McAb were shown by immunoblotting to react with the carboxy (C) terminal polypeptide of factor VIII which appeared as a doublet with a molecular weight (Mr) of 77,000/75,000. The C terminal factor VIII polypeptide was detectable by immunoblotting in each of 12 therapeutic factor VIII concentrates, from six different manufacturers, although its level was variable. Factor VIII was visualized in plasma by immunoblotting following its immunoadsorption and elution from agarose-bound monoclonal antibodies. No Mr 77,000/75,000 bands were detectable in plasma obtained from 13 unrelated CRM- haemophiliacs whilst 11 CRM+ haemophilic plasmas from seven kindred were shown to have a C terminal factor VIII polypeptide of normal molecular size.
已制备出两种针对人凝血因子VIII的单克隆抗体（McAb），分别命名为12A4和19C1。在免疫测定中，12A4与血浆中的凝血因子VIII抗原（VIII:Ag）结合，但不与血清中的结合，而19C1则与血浆和血清中的VIII:Ag均结合。免疫印迹显示，两种单克隆抗体均与凝血因子VIII的羧基（C）末端多肽发生反应，该多肽表现为分子量（Mr）为77,000/75,000的双峰。通过免疫印迹法可在来自六个不同厂家的12种治疗性凝血因子VIII浓缩物中检测到C末端凝血因子VIII多肽，尽管其水平各不相同。凝血因子VIII经免疫吸附并从琼脂糖结合的单克隆抗体上洗脱后，通过免疫印迹法在血浆中可见。在从13名无关的CRM - 血友病患者获得的血浆中未检测到分子量为77,000/75,000的条带，而来自七个家族的11份CRM + 血友病血浆显示具有正常分子大小的C末端凝血因子VIII多肽。
