中文摘要：

为制备鸡白介素10（chIL-10）单克隆抗体,以常规分子生物学技术从禽细胞系MDCC-MSB1克隆chIL-10基因,将去信号肽chIL-10基因亚克隆至原核表达载体p ET-30a,并在大肠杆菌中表达,纯化后作为免疫原免疫BALB/c小鼠。经4次免疫后,取小鼠脾细胞与骨髓瘤细胞（SP2/0）融合。将原核表达的GST-chIL-10融合蛋白作为包被抗原,利用间接ELISA筛选阳性克隆。阳性细胞株经3次亚克隆后,获得3株稳定分泌chIL-10抗体的杂交瘤细胞克隆,分别命名为4B2、4F3以及1E3。经间接ELISA测定,以上3株杂交瘤细胞腹水效价为1∶3.2×106,亲和力解离常数（Kd）分别为1.05×10-9、5.01×10-10以及7.59×10-10。抗体重链类型分别为Ig G2a、Ig G2a以及Ig G1。经Western Blot试验证明,3株单抗均能特异地识别原核或真核表达的chIL-10蛋白,4B2、4F3单抗识别的抗原表位区域为chIL-10 N端的1 aa52 aa,1E3识别的是N端的52 aa102 aa。这些单抗为鸡IL-10的检测及生物学功能研究奠定了基础。

英文摘要：

To develop monoclonal antibodies（ Mc Abs） against chIL-10,chicken interleukin-10（ mch-10） gene was cloned into a p ET- 30 a vector,and the protein was expressed in E. coli and purified with affinity Chromatography. We immunized BALB / c mice with the purified protein,and fused murine splenocytes with SP2 /0. An indirect ELISA using GST- mchIL-10 protein as coating antigen was established to screen positive clones. We obtained 3 hybridoma cell clones that stably secreted Mc Abs against chIL-10 and these clones were named 4B2,4F3 and 1E3,respectively. The titers in ascite fluid were 1： 3. 2 × 106. The dissociation constants（ k Ds） were 1. 05 × 10-9,5. 01 × 10-10,and 7. 59 × 10-10,respectively. The isotypes of these Mc Abs were determined to be Ig G2 a,Ig G2 a and Ig G1. These Mc Abs specifically bound to chIL-10 expressed by either prokaryotic or eukaryotic system as demonstrated by Western Blot. The binding domain of chIL-10 recognized by 4B2,4F3 was located between 1- 52 aa,and that by 4B2 was located between 52-102 aa as determined by Western Blot. These Mc Abs would help to detect chIL-10 and to elucidate the biological roles of chIL-10 in immune responses.