江苏大学学报（医学版）2025，Vol.35Issue(5)：376-380,5.DOI:10.13312/j.issn.1671-7783.y240113
TrkA-脂筏@琼脂糖凝胶色谱模型的构建及亲和行为研究
Construction of TrkA-lipid rafts@CNBr-Sepharose 4B gels chromatography and its affinity behavior
摘要
Abstract
Objective:To construct a bioaffinity chromatography of TrkA-lipid rafts@CNBr-Sepharose 4B gels and apply it to the assessment of the efficacy of the affinity adsorption of TrkA targeted drugs.Methods:TrkA-lipid rafts@CNBr-Sepharose 4B gels stationary phase was prepared and characterized through the coupling reaction of cyanogen bromide immobilized ligand.The stability was assessed by repeating the up-sampling experiment over a period of 30 days.Affinity adsorption experiments were conducted to investigate the affinity adsorption behavior of TrkA-targeted drugs on both blank agarose gels columns and TrkA-lipid rafts@CNBr-Sepharose 4B gels columns.Results:The Western blotting results revealed that the lipid rafts contained Flotillin-1,Caveolin-1,and TrkA proteins.Following the coupling of U251 cell lipid rafts coupled Sepharose 4B gels,it was observed that the lipid rafts maintained a stable solid loading on the column bed even after rinsing with multiple column volumes of buffer.Adsorption experiments confirmed the specific affinity adsorption of the TrkA-lipid rafts@CNBr-Sepharose 4B gels stationary phase.Conclusion:The bioaffinity chromatography of TrkA-lipid rafts@CNBr-Sepharose 4B gels was successfully constructed,thus preliminarily validating its feasibility and stability.关键词
原肌球蛋白受体激酶A/脂筏/琼脂糖凝胶/亲和吸附/吉非替尼Key words
TrkA/lipid rafts/CNBr-Sepharose 4B gels/affinity adsorption/gefitinib分类
医药卫生引用本文复制引用
沈鑫,池昊,唐唯佳,童珊珊..TrkA-脂筏@琼脂糖凝胶色谱模型的构建及亲和行为研究[J].江苏大学学报（医学版）,2025,35(5):376-380,5.基金项目
国家自然科学基金资助项目(81673574):镇江市重点研发计划(社会发展)项目(SH2023062) （81673574）
