摘要
应用ELISA定量结合试验和ELISA定量抑制试验,测定出三株鼠抗人B血型物质单克隆抗体3-3-D9(IgGl),3-5-D12(IgGl)和6-1-G11(IgA)结合部位的结构和特异性。研究发现三株抗B血型单克隆抗体仅结合B血型物质,而不结合A,H和L_e^a血型物质,表现出对B血型的高度特异性。同时这些单抗对B血型物质TijⅡ phenol-insoluble有较高亲和力,与另一B血型物质Beach phenol-insoluble亲和力较低,尽管Beach phenol-insoluble是6-1-G11免疫用抗原。3-3-D9和3-5-D12结合部位互补于含有双分子岩藻糖残基的。这一发现在世界上尚属首次。6-1-G11结合部位互补于含有单分子岩藻糖残基的02E.Glα1。这些单克隆抗体结合部位结构和特异性的揭示,为在临床血液分型试验中广泛应用这些抗B血型单克隆抗体,取代沿用已久的人多克隆抗血清奠定了基础。
The size of binding site and specificities of three mouse monoclonal anti-Bs:3-3-D9(IgGl) , 3-5-D12 (IgG1) and 6-l-Gll(IgA) ,have been characterized with quantitative ELISA binding assays and inhibition assays All of them bind to B blood group substances, but not to A, H and Lea blood group substances. All three monoclonal antibodies show higher affinity to Tij Ⅱ phenol-insoluble (B) than to Beach phenol-insoluble (B) B group substance, although Beach phenol -insoluble (B) was the antigen of 6-1-G11. It is found that 3-3-D9 and 3-5-D12 were most complementary to the difucosyl containing B-Penta. Galα1→3GaIβ1→4Glc,Fuca Fucathis is the first time that the existence of difucosyl containing structure on the B blood group determinants is reported in literature. The third monoclonal, 6-1-G11, was most complementary to the monofucosyl containing 02E: Galα1→3Galβ-O(CH2)8COOCH3. The establishment of the combining site and specifi-Fucacity of monoclonal anti-Bs is very important for using three monoclonals in clinical blood groups.
基金
国家教委优秀年轻教师基金
国家自然科学基金
关键词
单克隆抗体
结合部位
B血型物质
Antibody combining site
Monoclonal antibody
B blood group substance, ELISA
