Construction of and screening for phage antibody library of human lung adenocarcinoma
- VernacularTitle:人源肺腺癌噬菌体抗体库的构建及初步筛选
- Author:
Hua PANG
;
Yi LUO
;
Shaolin LI
- Publication Type:Journal Article
- Keywords:
phage antibody library;
single-chain variable fragment;
lung adenocarcinoma;
screening
- From:Journal of Third Military Medical University
2003;0(22):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct human single-chain antibody gene library associated with lung adenocarcinoma, and screen lung adenocarcinoma cell A549-specific antibody. Methods The lymphatic tissue near lung adenocarcinoma was used to construct human single-chain antibody gene library. The specific antibody to lung adenocarcinoma cell A549 was screened from the antibody library. Positive clone bacteria were transformed to E. coli HB2151, and their dissolvability was observed. The soluble scFv was purified by affinity chromatography and its relative molecular mass was determined by SDS-PAGE and Western blotting. The specificity of scFv to human lung adenocarcinoma cells was identified with ELISA. Results The phage antibody library of 4.6?107 was constructed successfully. Single-chain antibody of human lung adenocarcinoma was enriched. The ratio of yield was increased gradually, 181 times higher after 5 rounds of panning than after the first round of panning. SDS-PAGE and Western blotting results showed soluble scFv' molecular mass was about 30 000. ELISA showed dissolved antibody had high specificity to bind A549 cells, not MDA-MB-435. Conclusion The single-chain antibody gene library of human lung adenocarcinoma was constructed successfully, and specific antibodies to lung adenocarcinoma were screened, providing the basis for single-chain antibody radionuclide imaging and treatment.
