抗金玉兰酶制剂(β-内酰胺酶)单克隆抗体的研制与特性分析

Preparation and characterization of monoclonal antibodies against β-lactamase

以金玉兰酶制剂(β-内酰胺酶)为抗原,制备出16株能稳定分泌抗β-内酰胺酶特异单克隆抗体的杂交瘤细胞株。对其中的高效价单克隆抗体进行了特性和表位分析,结果显示,抗原金玉兰酶制剂分子表面至少有两个相同或相近的表位,且它们的距离较远,采用4F11、2 H5或1B10中的任一株单抗,就能实现对β-内酰胺酶的双抗夹心ELISA检测;4F11、2 H5和1B10的免疫球蛋白亚型鉴定都是IgG1,亲和力常数在1×107L/mol～1×1010L/mol之间;4F11的检测灵敏度为19 ng/mL。证实该3株单克隆抗体可用于研制检测牛奶中金玉兰酶制剂的免疫学试剂盒。

Using β-lactamase produced by Jin Yu-lan company as the antigen, 16 hyhridomas stably secreting McAbs to β-lactamase were developed. These McAbs with high antibody titers were detected on their specificities and epitopes. The results show that there are at least two epitopes which are the same or similar on the surface of the antigen and the distance between them are great. Therefore, β-lactamase could be detected by a two-antibody-sandwich enzyme-linked immunosorbent assay （das ELISA） formed by any McAb among 4F11, 2H5 and 1B1 0. 1 g subclasses of 4Fll, 2H5 and 1BIO are IgG1. Their affinity constants （Kaffi） are from 1 × 10^7 L/mol to 1 ×10^10 L/mol. The sensitivity of 4F11 in detectino of antigen is as low as 19 ng/mL. They can be used in developing the immunoassay kits for the detection of β-lactamase; hybridomas ; monoclonal antibody-lactamase in milk.