Dmitriev A D, Massino Iu S, Dergunova N N, Kizim E A, Segal O L, Smirnova M B, Vostrikov V M, Koliaskina G I
Vestn Ross Akad Med Nauk. 1996(4):46-51.
Bifunctional antibodies (bABs) having a double specificity to alpha-endorphin (alpha-END) and horseradish peroxidase (HRP) were produced by hybridoma technology. The antibodies constituted about 28-29% of all immunologically active IgG secreted by hybrid hybridoma (quadroma). The quadroma was isolated by fusion of two murine hybridomas (anti-HRP and anti-alpha-END) with distinct phenotypes: double mutant AMD(R)/NAT(S) and its wild type. To produce the double mutant phenotype, an actinomycin D-resistant (AMD(R)) mouse myeloma was used to initiate one of the parental hybridomas. bABs were purified from quadroma culture medium and ascitic fluids by sequential HRP-sepharose and alpha-END-sepharose affinity chromatography. With radioimmunoassay, the affinity of the individual anti-alpha-END combining sites of bABs was shown to be identical to that of parental monoclonal antibodies. Binding to the second antigen (HRP) did not affect the binding of bABs to alpha-END. bABs proved to be efficient for the determination of endorphins and their precursor proopiomelanocortin in immunohistology and immunoblotting.
利用杂交瘤技术制备了对α-内啡肽（α-END）和辣根过氧化物酶（HRP）具有双重特异性的双功能抗体（bABs）。这些抗体约占杂交杂交瘤（四瘤）分泌的所有免疫活性IgG的28%-29%。四瘤是通过将两种具有不同表型的鼠杂交瘤（抗HRP和抗α-END）：双突变体AMD(R)/NAT(S)及其野生型融合而分离得到的。为了产生双突变体表型，使用对放线菌素D耐药（AMD(R)）的小鼠骨髓瘤细胞来启动其中一个亲本杂交瘤。通过连续的HRP-琼脂糖和α-END-琼脂糖亲和层析从四瘤培养基和腹水中纯化bABs。通过放射免疫测定法表明，bABs的各个抗α-END结合位点的亲和力与亲本单克隆抗体的亲和力相同。与第二种抗原（HRP）的结合并不影响bABs与α-END的结合。在免疫组织化学和免疫印迹中，bABs被证明可有效地用于内啡肽及其前体阿片促黑皮质素的测定。
