人白细胞介素17的多克隆抗体制备与单抗杂交瘤的筛选

Preparation of Polyclonal Antibody of Recombinant Human Interleukin-17 and Screening of its Monoclonal Antibody Hybridoma

:目的 制备重组人IL - 17的兔抗血清并纯化 ,克隆化筛选IL - 17的阳性杂交瘤克隆。方法 应用纯化的IL - 17程序免疫家兔 ,得到兔抗人IL - 17的抗血清 ;采用盐析法和亲和层析法对兔抗血清进行纯化。应用纯化的IL - 17免疫小鼠 ,将其脾脏与骨髓瘤细胞SP 2 / 0融合 ,采用有限稀释法和克隆化筛选阳性杂交瘤克隆。抗体的检测分别采用琼脂糖双向扩散试验法和酶联免疫测定法 (ELISA)。结果和结论 纯化得到多克隆抗体 2 .5ml,IgG纯度大于 95 % ,初步得到 5个阳性杂交瘤克隆 。

Aim To prepare and purify of rabbit antiserum of recombinant human Interleukin-17(rhIL-17),and screen mouse positive hybridome clones of McAb by clone .Methods Antiserum was obtained by programmed immunization of rabbits with purified rhIL-17 and purified through ammonium sulfate precipitation and affinity chromatography.The spleen of immunized mouse was fused with myeloma cell line SP 2/0,and positive clones were obtained by definite dilution and cloned screening.Antibody was detected by double agar gel immune diffusion test and ELISA.Results and conclusion 2.5ml antiserum(polyclonal antibody) was purified which the purity of IgG was above 95%.5 positive clones were obtained which can secret monoclonal antibody.The antiserum and positive clones could be used in subsequent study of activity block,ELISA and affinity chromatograph purification of IL-17.