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目的:表达、纯化T7噬菌体衣壳蛋白P11,并制备其单克隆抗体(mAb)。方法:克隆并表达T7噬菌体P11蛋白,其氨基端带有6-His标签。纯化后的蛋白免疫BALB/c小鼠,经融合、筛选制备特异性mAb。结果:成功表达了P11蛋白。SDS-PAGE显示所表达蛋白的相对分子质量(Mr)约为27000。获得了1株稳定分泌抗P11抗体的杂交瘤细胞株(2G11),其分泌的mAb的Ig亚类(型)为IgG2b。ELISA检测,对应腹水mAb的效价为1∶8.1×105。Western blot结果显示抗P11mAb具有良好的特异性。结论:成功地制备了P11蛋白及其mAb。
Objective: To express and purify T7 phage coat protein P11 and prepare its monoclonal antibody (mAb). Methods: The P7 protein of T7 phage was cloned and expressed, and its 6-His tag was found at its amino terminus. BALB / c mice were immunized with the purified protein, and fused and screened to prepare specific mAb. Results: P11 protein was successfully expressed. SDS-PAGE showed that the expressed protein had a relative molecular mass (Mr) of about 27,000. A hybridoma cell line (2G11) stably secreting an anti-P11 antibody was obtained, and the Ig subclass (type) of the secreted mAb was IgG2b. ELISA, ascites mAb titers of 1: 8.1 × 105. Western blot results show that anti-P11 mAb has good specificity. Conclusion: P11 protein and its mAb were successfully prepared.