抗鸡IgM单克隆抗体的研究──Ⅲ．抗鸡IgMμ链单克隆抗体的制备及生物学特性鉴定

Resesarch on Monoclonal Antibodies Specific for Chicken IgM Ⅲ.Preparation of Monoclonal Antibodies Against μChain of Chicken IgM and Identification of Their Biological Characteristics

以亲和层析纯化的鸡ＩｇＭ免疫ＢＡＬＢ／Ｃ小鼠为材料，取３次免疫后的脾细胞与ＳＰ２／０骨髓瘤细胞融合，用ＥＬＩＳＡ筛选抗鸡ＩｇＭ和ＩｇＧ抗体以确定抗鸡ＩｇＭ重链特异性的抗体，获得了７株分泌抗鸡ＩｇＭμ链特异性单克隆抗体（ＭｃＡｂ）的杂交瘤细胞株．命名为Ｍ２、Ｍ４、Ｍ７、Ｍ９、Ｍ１０、Ｍ１１；和Ｍ１２。这７株杂交瘤细胞株连续传代６个月，并经液氮冻存５个月后复苏培养，仍能稳定地分泌特异性ＭｃＡｂ。７株ＭｃＡｂ均具有ＥＬＩＳＡ特性和免疫沉淀特性，腹水ＥＬＩＳＡ效价均在１０－５以上，其中Ｍ２、Ｍ４和Ｍ１０３株腹水效价高达１０－１０以上，最高者达到１０－１５。琼脂免疫扩散试验和ＥＬＩＳＡ交叉反应性试验测定表明，７株ＭｃＡｂ均与鸡ＩｇＭ呈阳性反应，与鸡ＩｇＧ呈阴性反应。用羊抗鸡ＩｇＭμ链抗血清进行ＥＬＩＳＡ阻断试验，结果表明，７株ＭｃＡｂ均是抗鸡ＩｇＭμ链的ＭｃＡｂ。

Seven hybridoma cell lines (desingated M2, M7, M9, M10, M11 and M12) Were estsblishsd Which secreted antibodies specific for chicken IgM by fusion of SP2/0 myelome cells with splenocytes from BALB/C mice triimmunized with purified chicken IgM by affinity chromatogra phy from chicken serum. The specificity for the IgM heavy chain was shown by using ELISAs to screen for antibodies to IgM and IgG. The monoclonal antibodies(McAb) specific for chicken IgM could be secreted steadily by all of these lines after six months' serial passages and five months' preservation in liquid nitrogen. All the McAbs had both the properties of ELISA and immunoprecipitability. The titers of ELISA of these McAbs in ascites were all above 1∶105; those of M2, M4 and M10 were above 1∶1010;and the highest titer reached 1∶1015. All of these McAbs reacted only with chicken IgM, but not with chicken IgG byagar immunodiffusion test and cross-reactivity test with ELISA. Furthermore, all the McAbs were specific for μchain of chick en IgM in ELISA block test with serum of goat anti-μ chain of chicken IgM.