[Preparation and identification of monoclonal antibodies against human pulmonary surfactant-associated protein A].
OBJECTIVE
To prepare monoclonal antibody (mAb) against human pulmonary surfactant-associated protein A (SP-A) and evaluate its specificities.


METHODS
The hybridoma cell line secreting SP-A mAb was established to induce the production of SP-A ascites fluid in BALB/c mice immunized with SP-A. The ascites fluid was then collected and purified through (NH4)2SO4 salting-out procedure, and the titer and specificity of the purified monoclonal anti-SP-A antibodies were determined by Western blotting and indirect enzyme-linked immunosorbent assay, respectively.


RESULTS
Three hybridoma cell lines capable of specific antibody secretion were established, among which 2B6 produced the antibody of the highest titer, with the hybridoma cell chromosome exceeding 100. The antibody titers in the ascitic fluid and cell supernatant fluid were 1: 50,000 and 1: 10,000 respectively, and Western blotting displayed a reaction band representing a relative molecular mass of 31,000, which was produced by the monoclonal antibody in reaction to purified SP-A.


CONCLUSIONS
The antibodies of high purity and specificity against human SP-A have been successfully prepared.
Liu
Yong-mei
YM
Department of Pediatrics, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China.
Feng
Zhi-chun
ZC
Du
Jiang
J
Wu
Bing-yi
BY
Tan
Yue-er
YE
Wang
Xian-yan
XY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D037662:Pulmonary Surfactant-Associated Protein A / Q000276:immunology*
D000911:Antibodies, Monoclonal; D037662:Pulmonary Surfactant-Associated Protein A