【摘要】 目的:制备和初步鉴定抗结核分枝杆菌抗原优势肽段的单克隆抗体。方法:以两种优势肽段的融合抗原PGEX-4T-2/38kD-ESAT6-CFP10和PGEX-4T-2/Mtb8.4-MPT64-Mtb16.3-Mtb8作为免疫原免疫BALB/c小鼠,取其脾细胞与SP2/0小鼠骨髓瘤细胞融合后,通过有限稀释法筛选阳性克隆,经ELISA和Western blotting分析单抗的特性和特异性。结果:获得17株结核分枝杆菌抗原优势肽段特异单克隆抗体杂交瘤细胞株,经间接ELISA方法测定,杂交瘤细胞培养上清效价为28～212,接种小鼠的腹水效价为216～220。Western blotting结果显示,每株单抗均能特异性的识别融合蛋白的优势肽段。结论:制备了针对抗结核分枝杆菌抗原优势肽段的单克隆抗体,为结核分枝杆菌的快速诊断和抗原性分析奠定了基础。更多还原

【Abstract】 Objective:To develop and characterize monoclonal antibodies(McAbs) against the dominant peptide antigen of Mycobacterium tuberculosis.Methods: BALB/C mice were immunized with two fused dominant peptide antigen(PGEX-4T-2/38kD-ESAT6-CFP10,PGEX-4T-2/ Mtb8.4-MPT64-Mtb16.3-Mtb8) of Mycobacterium tuberculosis.Splenocytes from the immunized mice were fused with SP2/0 myeloma cells.Hybridoma culture supernatants were screened and evaluated by ELISA.Positive clones were checked for isotype and McAbs were confirmed by Western blotting analysis.Results: Seventeen strains of stable hybridoma cells secreting Mcab against the fused dominant peptide antigen of Mycobacterium tuberculosis were obtained.The titers of these Mcabs in the culture supernatants were 28-212,and the tietrs in the ascites of the mice were 216-220 detected by ELISA.The results of western blotting showed that each Mcab only reacted with the dominant peptide of the fused antigen.Conclusion: McAbs specific to fused dominant peptide antigen of Mycobacterium tuberculosis were developed.McAbs would be helpful for rapid clinical diagnosis and antigenic epitope analysis for Mycobacterium tuberculosis.更多还原