Tan Feng, Pan Chang-wang, Liiang Shao-hui, Huang Hui-cong
Department of Parasitology, Wenzhou Medical College, Wenzhou 325035, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2005 Aug 30;23(4):209-12.
To develop and identify monoclonal antibodies (McAbs) against adult worm of Angiostrongylus cantonensis and observe its applicability.
BALB/c mice were immunized with soluble antigen of adult worms of A. cantonensis. The spleen cells of immunized mice were fused with myeloma cell, and the hybridoma secreting high titer of McAbs with high specificity was screened. By using the McAbs, serum of angiostrongyliasis patient and sera of the rats infected with A. cantonensis were detected by Western blotting and double antibody sandwich ELISA respectively.
Three McAbs were established (2A2, 3F1, 4H2), which all showed no cross reaction with antigens of Schistosoma japonicum, Paragonimus westermani, Cysticercus cellulosae and Trichinella spiralis. Western blotting analysis demonstrated that the three McAbs recognized a Mr 15,000 soluble antigen of adult worm of A. cantonensis and recognized the Mr 24,000 and Mr 15,000 circulating antigens from the serum of angiostrongyliasis patient. The double antibody sandwich ELISA detection showed a positive rate of 76.5%.
Three hybridoma cell lines against adult worm of A. cantonensis have been established which secret high titer of McAbs with high specificity and seem promising in detecting the circulating antigen of the angiostrongyliasis patient.
制备并鉴定抗广州管圆线虫成虫的单克隆抗体（McAbs），并观察其适用性。
用广州管圆线虫成虫可溶性抗原免疫BALB/c小鼠。将免疫小鼠的脾细胞与骨髓瘤细胞融合，筛选分泌高特异性、高滴度McAbs的杂交瘤。分别用Western印迹法和双抗体夹心ELISA法，使用所制备的McAbs检测广州管圆线虫病患者血清及感染广州管圆线虫大鼠的血清。
建立了3株单克隆抗体（2A2、3F1、4H2），它们与日本血吸虫、卫氏并殖吸虫、猪囊尾蚴和旋毛虫抗原均无交叉反应。Western印迹分析表明，这3株单克隆抗体识别广州管圆线虫成虫的一种相对分子质量为15 000的可溶性抗原，且能识别广州管圆线虫病患者血清中相对分子质量为24 000和15 000的循环抗原。双抗体夹心ELISA检测阳性率为76.5%。
已建立3株抗广州管圆线虫成虫的杂交瘤细胞系，它们分泌高特异性、高滴度的单克隆抗体，在检测广州管圆线虫病患者循环抗原方面具有良好的应用前景。
