摘 要：
采用饱和硫酸铵盐析、离子交换柱层析技术从猪血清中提纯IgG.经SDS-PAGE电泳鉴定,纯化的IgG达到电泳纯.以此纯化的IgG,采用长程免疫法免疫BALB/c小鼠,取4次免疫后的脾细胞与SP2/0骨髓瘤细胞用PEG-1500进行细胞融合,融合率为90.3%.建立间接ELISA,用于检测杂交瘤细胞上清中的单克隆抗体.初检阳性率为13.9%.经2次亚克隆反复筛选获得了2株能稳定分泌抗猪IgG单克隆抗体的杂交瘤细胞株,命名为6B4、4B8.ELISA证实所获得的单抗是特异性针对IgG,6B4、4B8经反复冻存,复苏及多次传代,仍能分泌高效价抗体,分泌抗体均属IgG1、κ型.本研究所获得的单克隆抗体将对检测猪抗体水平及IgG提纯发挥重要作用.
译 名：
Preparation of Monoclonal Antibodies Against Porcine IgG
作 者：
WU Sheng-xi,CAI Jia-li,HU Ren-jian Bioengineering College,Chongqing Institute of Technology,Chongqing 400050,China
关键词：
chromatography;porcine IgG;monoclonal antibodies
摘 要：
Porcine immunoglobulin G(IgG)was isolated and purified from porcine serum by a procedure of precipitation with saturated ammonium sulfate and ion-exchange chromatography with DEAE52,and was identified by SDS-PAGE electrophoresis.BALB/c mice were immunized four times with the purified IgG,then the spleen cells of the hyperimmunized mice and SP2/0 cells were fused with PEG-1500.A cell fusion rate of 90.3% was obtained.An indirect enzyme-linked immunosorbent assay(ELISA)was developed to detect the monoclonal antibodies(Mabs)secreted by the hybridoma cell lines and a positive rate of 13.9% was obtained.Two hybridoma cell lines secreting monoclonal antibodies,named 4B8 and 6B4,were developed by two subclones.The specificity of Mabs against IgG was identified by means of indirect ELISA.After repeated frozen storage,thawed revival and generation,they kept steadily producing high titer of Mabs.The two McAbs both had mouse IgG1,κ isotype.The Mabs prepared would be very useful in detecting the level of IgG and purifying IgG.
