中文摘要：

目的构建人源肺腺癌噬菌体单链抗体库，并筛选肺腺癌细胞A549特异性单链抗体。方法利用肺腺癌患者癌旁淋巴结组织构建肺腺癌噬菌体单链抗体库。从该抗体库中筛选特异性识别A549细胞的单链抗体，将阳性克隆菌转化E．coliHB2151进行可溶性表达。抗体亲和层析纯化后经SDS—PAGE、Westernblot鉴定，通过ELISA法鉴定其与人肺腺癌细胞结合的特异性。结果成功构建了1个4．6×10^7的噬菌体抗体库。在筛选过程中，肺腺癌噬菌体单链抗体得到富集，收获率逐轮得到提高，第5轮为第1轮的181倍。在E．coliHB2151中实现了单链抗体的可溶性表达。SDS—PAGE与Westernblot结果显示抗体相对分子质量为30×10^3左右。细胞ELISA测定结果显示可溶性抗体具有较高的特异性，能与A549细胞结合，而不与MDA—MB-435细胞结合。结论从肺腺癌病人癌肿周围淋巴结扩增免疫球蛋白基因成功地构建人源肺腺癌噬菌体单链抗体库，并从中筛选到肺腺癌特异性抗体。

英文摘要：

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 × 10^7 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.