高等学校化学学报Issue(12)：2551-2555,5.DOI:10.7503/cjcu20140421
靶向喉癌的蜂毒肽重组抗体的构建及鉴定
Synthesis and Characterization of a Melttin Expressing Recombinant Antibody Targeting Human Laryngeal Cancer
摘要
Abstract
Laryngeal carcinoma is poor prognosis and patients with laryngeal carcinoma usually present late leading to the reduced treatment efficacy and high rate of recurrence. Incidence and mortality rates of laryngeal carcinoma are equivalent, suggesting a failure of current therapies, despite the variety of combined modality approaches which have been introduced to complement surgical treatment. A successful cancer therapeutic should target tumours specifically with limited systemic toxicity. Herein, the recombinant antiEGFR/MEL pro-tein was expressed in Escherichia coli( E. coli) , refolded and purified on an immobilized Ni2+-affinity chroma-tography column. In the protein, anti epidermal growth factor receptor( EGFR) single chain antibody was used to target the EGFR in the laryngeal cancer cell, and the melittin was used to mediate inhibition of cell growth. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis( SDS-PAGE) and Western blotting analysis revealed that antiEGFR/MEL was sufficiently expressed. Confocal microscopy and flow cytometry demonstrated that antiEGFR/MEL bound specifically to Hep-2 cells, as almost no binding to Jurkat cells was observed under identical time and dosage conditions. MTT assay showed that antiEGFR/MEL suppressed the growth of Hep-2 cells effecitively. Collectively, these results suggest that antiEGFR/MEL is biologically active and specific toward EGFR-positive tumor cells and may represent an effective EGFR-targeted cancer therapy.关键词
喉癌/表皮生长因子受体/单链抗体/蜂毒肽/原核表达Key words
Laryngeal carcinoma/Epidermal growth factor receptor/Single chain antibody/Melittin/Prokaryotic expression分类
化学化工引用本文复制引用
孙丽丽,王浩然,徐一鸣,费丹,付婷婷,管国芳..靶向喉癌的蜂毒肽重组抗体的构建及鉴定[J].高等学校化学学报,2014,(12):2551-2555,5.基金项目
国家自然科学基金(批准号:81101140)、国家“八六三”计划项目(批准号:2012AA02A407)和吉林省科技发展计划重点项目(批准号:20130206041NY)资助.覮 Supported by the National Natural Science Foundation of China(No.81101140), the National High Technology Research and Development Program of China ( No.2012AA02A407) and the Key Technologies Research and Development Program of Jilin Province, China ( No.20130206041NY) （批准号:81101140）
