人肝癌细胞系HLE细胞表面抗原多肽的提取、纯化以及初步鉴定

The extraction, purification and identification of antigen-derived peptides expressed on the surface of HLE cells

目的 提取人肝癌细胞系HLE细胞表面抗原多肽并进行初步鉴定。方法 应用弱酸洗涤法从HLE细胞表面洗脱出与MHC I类分子结合的抗原多肽,经低温冷冻干燥浓缩、反相高效液相色谱仪(RP HPLC)纯化后,用 Lowry法进行定量并以基质辅助激光解吸电离 飞行时间质谱仪(MALDI TOF MS)对其成分进行初步鉴定。结果 应用弱酸洗涤法可从每109 个HLE细胞表面提取抗原多肽(52 3±8 6)μg,经HPLC及MALDI TOF MS鉴定其中有 31 种多肽组分。结论 使用弱酸洗涤法、低温冷冻干燥以及HPLC方法可以从肝癌细胞表面洗脱并纯化出多种与MHC I类分子结合的抗原多肽。

Objective To extract, purify and identify the antigen derived peptides bound to MHC class I molecules on the cell surface of human hepatocellular carcinoma cell line HLE Methods Antigen peptides were extracted by mild acid elution, concentrated by lyophilization, and fractioned by reversed phase high performance liquid chromatography (RP HPLC) Lowry assay was performed for the quantitation of purified sample and the peptides were identified by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI TOF MS) Results (52 3±8 6)μg antigen derived peptides could be extracted from the surface of 1×10 9 HLE cells by mild acid elution, 31 kinds of components contained in the elution peptide samples were identified by HPLC and MALDI TOF MS Conclusion Antigen derived peptides bound to MHC class I molecules on the cell surface of human hepatocellular carcinoma could be extracted and purified by mild acid elution, lyophilization and HPLC