摘要
采用重氮化法将SM2分别与BSA、OVA偶联,合成人工免疫抗原SM2-BSA和人工检测抗原SM2-OVA,并通过紫外光谱扫描(UV)、SDS-PAGE、红外光谱扫描(IR)验证偶联成功,偶联比分别为14∶1、10.5∶1。用SM2-BSA免疫BALB/c小鼠,通过细胞融合技术制备了2株特异性分泌SM2抗体的杂交瘤细胞株,分别是3A12和4H4,亲和力常数均为1×108～1×1010 L/mol,3A12的检测灵敏度为0.6 ng/mL,免疫球蛋白亚型鉴定为IgG1,4H4的检测灵敏度为1.8 ng/mL,免疫球蛋白亚型鉴定为IgG2α,这两株抗体均与其他6种磺胺类无交叉反应。
Sulfamethazine（SM2） was bound to bovine serum albumin（BSA） and ovalbumin（OVA） respectively by using diazotization,which aimed to synthesize artificial immunogenic antigen SM2-BSA and artificial detection antigen SM2-OVA.The results of UV scan,SDS-PAGE,IR scan indicated that the artificial immunogenic antigen SM2-BSA and the artificial detection antigen SM2-OVA were synthesized successfully and their conjugation ratio was 14∶1 and 10.5∶1,respectively.BALB/c mice were immunized with SM2-BSA,and two hybridoma strains（named 3A12 and 4H4,respectively） which could specifically secrete antibody of SM2 were obtained by using cell fusion technique.The affinity constants（Kaffi） of 3A12 and 4H4 were between 1×108 L/mol and 1×1010 L/mol;the detection sensitivity of 3A12 was 0.6 ng/mL and its subtype of immunoglobulin was IgG1;the detection sensitivity of 4H4 was 1.8 ng/mL and its subtype of immunoglobulin was IgG2a.There was no cross-reactivity between the both antibodies and the other six species of sulfonamides.
出处
《湖南农业科学》
2011年第7期108-111,共4页
Hunan Agricultural Sciences
基金
河北省科技支撑计划项目(09227134D)
