Pimm M V, Robins R A, Embleton M J, Jacobs E, Markham A J, Charleston A, Baldwin R W
Cancer Research Campaign Laboratories, University of Nottingham, UK.
Br J Cancer. 1990 Apr;61(4):508-13. doi: 10.1038/bjc.1990.115.
A bispecific monoclonal antibody, reactive with methotrexate (MTX) and a tumour associated antigen (gp72) has been produced by fusing spleen cells from MTX immunised mice with 791T/36/3 (anti-gp72) hybridoma. The hybrid antibody was purified from anti-MTX and anti-gp72 antibodies present in the hybridoma culture supernatant by combinations of affinity chromatography on a MTX-agarose immunoabsorbent and stepwise acid elution from Sepharose-Protein A. A particular feature of the present antibody is that it reacts with conjugated MTX; this would allow in vivo targeting of conjugates, increasing many fold the number of molecules of drug carried or localising to pre-targeted antibody. Dual binding between tumour cell surface antigen and MTX was demonstrated by the ability of the hybrid antibody to bridge between tumour cells and MTX as MTX-HSA conjugate, reaction here being detected by flow cytofluorimetry. Purified hybrid antibody specifically enhanced the in vitro cytotoxicity of MTX-HSA for gp72 positive tumour cells.
通过将甲氨蝶呤（MTX）免疫小鼠的脾细胞与791T/36/3（抗gp72）杂交瘤融合，制备了一种与MTX和肿瘤相关抗原（gp72）反应的双特异性单克隆抗体。通过在MTX - 琼脂糖免疫吸附剂上进行亲和层析以及从琼脂糖 - 蛋白A上逐步酸洗脱，从杂交瘤培养上清液中存在的抗MTX和抗gp72抗体中纯化出杂交抗体。本抗体的一个特别之处在于它能与偶联的MTX反应；这将允许体内靶向偶联物，使携带的药物分子数量增加许多倍或定位于预先靶向的抗体。杂交抗体作为MTX - HSA偶联物在肿瘤细胞和MTX之间形成桥联的能力证明了肿瘤细胞表面抗原与MTX之间的双重结合，此处的反应通过流式细胞荧光术检测。纯化的杂交抗体特异性增强了MTX - HSA对gp72阳性肿瘤细胞的体外细胞毒性。
