發明
中華民國
110132354
I 788986
位向性共價鍵醣蛋白固定方法及抗體晶片
國立清華大學
2023/01/01
本發明提供一種位向性共價鍵醣蛋白固定方法及抗體晶片。固定方法包括提供配置有炔基化表面之氧化亞銅奈米粒子及銀塗佈固體表面。藉由表面氧化亞銅奈米粒子自催化疊氮-炔環加成反應將含疊氮基之硼酸甲苯磺醯探針固化至氧化亞銅奈米粒子-銀塗佈固體表面。提供醣蛋白至硼酸甲苯磺醯探針,接著藉由醣蛋白上的醣體或抗體Fc片段上醣鏈的醇基與硼酸甲苯磺醯探針的硼酸基形成有機硼酸酯, 將醣蛋白或抗體抓取於固體表面。經醣蛋白或抗體上鄰近探針的親核殘基經雙分子親核取代反應取代甲苯磺醯基,以透過共價鍵生成方式將醣蛋白固定於表面。 The invention provides an oriented and covalent method for immobilizing a glycoprotein or an antibody on a chip. The method includes providing a silver-coated solid surface equipped with alkynes and cuprous oxide nanoparticles. The azido boronic acid tosyl probe is conjugated to the silver-coated solid surface by the cuprous oxide nanoparticles through the self-catalyzed azide−alkyne cycloaddition. The glycan(s) of a glycoprotein or an antibody is provided to the boronic acid tosyl probe, and alcohol groups of the glycan(s) of the glycoprotein or the antibody and the boronic acid group of boronic acid tosyl probe form boronate ester. The nucleophilic residues on the glycoprotein or the antibody replace the tosyl group by SN2 reaction, so as to immobilize the glycoprotein or the antibody through the covalent bond formation.
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