Bedi G S, Back N
Adv Exp Med Biol. 1986;198 Pt A:127-35. doi: 10.1007/978-1-4684-5143-6_18.
Spleen cells from Balb/c mice immunized with purified rat plasma kininogen were fused to P-3 mouse myeloma cells. Positive clones were identified by enzyme linked immunosorbent assay (ELISA), cloned successively two times with limiting dilution and expanded as ascites tumors. Five hybridomas were developed that produced monoclonal antibodies against plasma kininogen. Two of the secreted antibodies were of the IgG1 (k) isotype and the remaining three were of the IgG1(lambda), IgG2A(k) and IgM(k) isotypes respectively. The specificity of the monoclonal antibodies was confirmed by the immunoprecipitation of kininogen with the antibodies coupled to Sepharose-4B followed by SDS-polyacrylamide gel electrophoresis. These monoclonal antibodies recognize at least two distinct epitopes on rat plasma kininogen.
用纯化的大鼠血浆激肽原免疫的Balb/c小鼠的脾细胞与P-3小鼠骨髓瘤细胞融合。通过酶联免疫吸附测定(ELISA)鉴定阳性克隆,用有限稀释法连续克隆两次并作为腹水肿瘤进行扩增。获得了五个产生抗血浆激肽原单克隆抗体的杂交瘤。分泌的抗体中有两种是IgG1(κ)同种型,其余三种分别是IgG1(λ)、IgG2A(κ)和IgM(κ)同种型。通过将抗体与Sepharose-4B偶联后对激肽原进行免疫沉淀,然后进行SDS-聚丙烯酰胺凝胶电泳,证实了单克隆抗体的特异性。这些单克隆抗体识别大鼠血浆激肽原上至少两个不同的表位。