抗幽门螺杆菌中性粒细胞激活蛋白的单克隆抗体的制备和鉴定
幽门螺杆菌;中性粒细胞激活蛋白;单克隆抗体;Westernblot;免疫组化,龙敏,罗军,李妍,李明,通讯作者:,PreparationandidentificationofmonoclonalantibodiesagainstHeli
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龙敏, 罗军, 南方医科大学公共卫生与热带医学学院微生物系 广东省广州市 5105155
李妍, 李明, 南方医科大学生物技术学院 广东省广州市 510515
广东省自然科学基金资助项目, No. 5004750
通讯作者: 李明, 510515, 广东省广州市, 南方医科大学生物技术学院. limin@fimmu.com
电话: 020-61648550
收稿日期: 2007-01-22 接受日期: 2007-03-06
Preparation and identification of monoclonal antibodies against Helicobacter pylori neutrophils activating protein
Min Long, Yan Li, Jun Luo, Min Li
Min Long, Jun Luo,Department of Microbiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, Guangdong Province, China
Yan Li, Min Li,School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong Province, China
Supported bythe Natural Science Foundation of Guangdong Province, No. 5004750
Correspondence to:Min Li, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong Province, China. limin@fimmu.com
Received:2007-01-22 Accepted:2007-03-06
Abstract
AIM: To screen and identify the monoclonal antibodies (mAbs) against H pylori neutrophils activating protein (NAP).
METHODS: H pylori DY01 and DY04 strains were clinically isolated. After BALB/c mice were immunized, the mAbs against H pylori were obtained by hybridoma technique. Then, the mAbs against H pylori-NAP were screened using recombinant H pylori-NAP. The obtained mAbs were evaluated for subtype and titer, and their specificities were identified by Western blot and immunohistochemical techniques.
RESULTS: Three strains of mAbs against H pylori-NAP were obtained. All of the mAbs against H pylori-NAP were identified as immunoglobulin G1 (IgG1) with k light chain. The titer of mAbs in the culture supernatant was 1/16 to1/32, and that in the ascites were 1/32000 to 1/64000. Western blotting showed that the mAbs had specific reaction with the recombinant H pylori-NAP. Clinical H pylori strains were stained brown with the mAbs against H pylori-NAP by immunohistochemical techniques ......
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