[Preparation and characterization of monoclonal antibodies against human secretory leukocyte protease inhibitor].
OBJECTIVE
To prepare and identify monoclonal antibodies(mAbs) against human secretory leukocyte protease inhibitor (hSLPI).


METHODS
BALB/c mice were immunized with hSLPI, and hybridoma cell lines were obtained by fusing mouse spleen cells with myeloma NS-1 cells. The specificity of mAbs were characterized by ELISA, Western blot, immunohistochemical staining, flow cytometry(FCM) and confocal laser scanning microscopy(CLSM).


RESULTS
Four hybridoma cells which secreted the mAbs to hSLPI were obtained. 4 mAbs were IgM. Western blot analysis showed that the mAbs could recognize a target molecule with relative molecular mass of 12 000. Immunohistochemical staining revealed that the reactivities of 4 mAbs to the epithelial cells in lung and colon tissues, mast cell-like cells in lung, colon, tonsil and prepuce tissues were positive. The result of FCM showed that the 4 mAbs recognized SLPI expressed in A549 cells. CLSM examination confirmed that the fluorescent markers were mainly localized in the cytoplasm of A549 cells.


CONCLUSIONS
The mAbs against hSLPI are prepared successfully, which provides valuable tool for studies on allergic and inflammatory diseases.
Chen
Yu-zhen
YZ
Research Institute of Allergy and Inflammation, Shantou University Medical College, Shantou 515031, China. g_yzchen@stu.edu.cn
He
Shao-heng
SH
Zhou
Yan-chun
YC
Huang
Tian
T
Liu
Yue-ning
YN
Chen
Lin-fei
LF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D053481:Secretory Leukocyte Peptidase Inhibitor / Q000276:immunology*
D000911:Antibodies, Monoclonal; D053481:Secretory Leukocyte Peptidase Inhibitor