摘要
〔目的〕 为建立岩沙海葵毒素(PTX)的酶联免疫快速测试方法提供特异性免疫抗体。〔方法〕 巯基化标准PTX成半抗原PTX -PDP ;巯基化匙眼血蓝蛋白(KLH)成KLH -SH ,两者合成反应人工抗原PTX -PDP -KLH。用PTX -PDP -KLH免疫雌性6～8周龄Balb/c小鼠,制备PTX多克隆抗体。用同样方法制备包被抗原PTX-PDP -BSA ,用以检测血清抗体效价。〔结果〕 紫外检测标准品PTX的最大吸收波长在2 6 4nm处,纯品KLH及BSA的最大吸收波长在2 78nm及2 79nm处,小鼠血清多克隆抗体PTX -PDP -KLH和PTX -PDP -BSA的最大吸收波长分别在2 6 7nm和2 73nm处。抗血清效价达到1:32 0 0 ,与正常Balb/c小鼠、昆明小鼠及兔血清间无交叉反应。〔结论〕 合成的PTX人工抗原纯度高,成功制备PTX多克隆抗体血清。
Objective:To produce the conjugates of keyhole limpet hemocyanin and chemically defined palytoxin haptens. Method: Palytoxin haptens were produced by derivatization of the primary amino group with succinimidyl 3-(2-pyridyldithio)propionate. Keyhole limpet hemocyanin were thiolated by 2-iminothiolane. Artificial antigens which were synthesized conjugating PTX-PDP to KLH-SH were used to immunized Balb/c mice to obtain polyclonal antibodies. PTX-PDP-BSA was produced through the same method and was used to of serum antibody titers. Results: According to the pictures of ultraviolet probe: the maximum absorbtion wave length of PTX was in 264nm and KLH in 278nm. But the maximum absorbtion wave lengths of PTX-PDP-KLH and PTX-PDP-BSA were in 267nm and 273nm respectively. The serum antibody titers reached to 1:3200. Conclusion: The antigen is very effective . The research provides the preparation for producing McAb of PTX and preparation the ELISA method of detecting PTX.
出处
《检验检疫科学》
2005年第2期3-6,共4页
Inspection and Quarantine Science
基金
国家科技攻关计划课题 (2 0 0 1BA80 4A2 0 - 2 - 4 )
