重组人γ-型干扰素单克隆抗体的鉴定

5株小鼠杂交瘤细胞ID5、1C10、1C11、3H9和6F8，所分泌抗重组人γ－型干扰素（rIFN－γ）单克隆抗体经双扩散试验证明其中4种为小鼠IgG1型，另一种为IgG2b型。用ELISA间接法测定小鼠腹水抗体效价为1：8000～1：128000。经免疫印迹试验证实5种单抗均能特异地识别rIFN－γ，其中1D5、3H9和1C11单抗对rIFN－γ具有中和活性，中和效价＞1：8000，用2种单抗建立的ELISA夹心法检测rIFN－γ，最低可测值为10．0ng／ml，且与白细胞介素－2，重组人α和β干扰素无交叉反应。

Characterization of McAb to rIFN-γ

The anti-rIFN-γ McAbs secreted by 5 strains of murine hybridoma cell (1D5,1C10, 1C11, 3H9 and 6F8) were identified by double immunodiffusion. The results proved that the McAbs secreted by 4 of the 5 strains were IgG1, and that secreted by the rest one was IgG2b. The antibody titers in murine ascites detected by indirect ELISA were 1: 8000~ 1: 128000. Western blot demonstrated that the McAbs secreted by all the 5 strains could recognize rIFN-γ specifically. Of the McAbs, those secreted by 1D5, 3H9 and 1C11 showed neutralizing activities to rIFN-γ, and the neutralizing titers of them were above 1: 8000.The sandwich ELISA method established using the McAbs secreted by 3H9 and 1C11 was used to detect rIFN-γ. The result showed that lowest detection limit was 10.0ng/ml, and no cross reaction with IL - 2, rIFN -α or rIFN -β was found.