Morris Todd A, Peterson Alexander W, Tarlov Michael J
National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, Maryland 20899, USA.
Anal Chem. 2009 Jul 1;81(13):5413-20. doi: 10.1021/ac900715d.
Glycoanalysis is important in the manufacture and quality control of protein therapeutics. An emerging method for glycoanalysis is the use of lectin arrays. Critical to the performance of these arrays is the immobilization of lectin molecules. Polydopamine has recently been shown to adsorb to a wide variety of surfaces. In this study, polydopamine (pDA) was used to modify gold, indium, and iridium surfaces and promote the adhesion of the alpha-mannose-specific lectin concanavalin A (Con A). The activity of the surface-bound lectin was demonstrated with the alpha-mannose-presenting glycoprotein ribonuclease B (RNase B). Surface plasmon resonance spectroscopy (SPRS) was used to demonstrate the selective affinity of RNase B for Con A. Surface-MALDI-TOF MS experiments revealed that the affinity of polydopamine-immobilized Con A for the glycoforms of RNase B is significantly affected by slight variations in oligosaccharide structure and composition. Specifically, surface-bound Con A binds certain Man7, Man8, and Man9 RNase B glycoforms more strongly than Man5 and Man6 glycoforms.
糖基分析在蛋白质治疗药物的生产和质量控制中具有重要意义。一种新兴的糖基分析方法是使用凝集素芯片。这些芯片性能的关键在于凝集素分子的固定化。最近研究表明，聚多巴胺能吸附到多种表面。在本研究中，聚多巴胺（pDA）用于修饰金、铟和铱表面，并促进α-甘露糖特异性凝集素伴刀豆球蛋白A（Con A）的附着。通过呈现α-甘露糖的糖蛋白核糖核酸酶B（RNase B）证明了表面结合凝集素的活性。表面等离子体共振光谱（SPRS）用于证明RNase B对Con A的选择性亲和力。表面基质辅助激光解吸电离飞行时间质谱（Surface-MALDI-TOF MS）实验表明，聚多巴胺固定的Con A对RNase B糖型的亲和力受寡糖结构和组成的微小变化显著影响。具体而言，表面结合的Con A对某些Man7、Man8和Man9 RNase B糖型的结合比Man5和Man6糖型更强。
