Schnaar R L, Lee Y C
Biochemistry. 1975 Apr 8;14(7):1535-41. doi: 10.1021/bi00678a030.
A new and versatile method for linking biologically active ligands to a polyacrylamide matrix is reported. Active esters of acrylic acid (N-succinimicyl acrylate and N-phthalimidyl acrylate) were synthesized, then copolymerized with acrylamide and N,N'-methylenebisacrylamide. Displacement of the active ester in the gel thus formed by various ligands containing aliphatic amino groups resulted in the formation of stable amid bonds between the ligands and the polyacrylamide gel. The affinity gel thus prepared has the following advantages: (i) resistance to chemical and microbiological degradation, (ii) ease of control of ligand level and higher levels of ligand possible, (iii) ease of control of porosity, and (iv) total displacement of the active ester under suitable conditions. Efficacy of this system was tested by preparation of 6-aminohexyl 2-acetamido-2-deoxy-beta-D-glucopyranoside derivative polyacrylamide gel by the described method. It was found to be more effective for purification of wheat germ agglutinin than the previously published affinity chromatography systems and the wheat germ hemagglutinin was obtained in crystalline form. In addition, partial resolution of isolectins was obtained from the affinity gel witha pH gradient.
报道了一种将生物活性配体连接到聚丙烯酰胺基质上的新型通用方法。合成了丙烯酸的活性酯（N-琥珀酰亚胺基丙烯酸酯和N-邻苯二甲酰亚胺基丙烯酸酯），然后与丙烯酰胺和N，N'-亚甲基双丙烯酰胺共聚。由含脂肪族氨基的各种配体取代由此形成的凝胶中的活性酯，导致配体与聚丙烯酰胺凝胶之间形成稳定的酰胺键。如此制备的亲和凝胶具有以下优点：（i）耐化学和微生物降解，（ii）易于控制配体水平且可能达到更高的配体水平，（iii）易于控制孔隙率，以及（iv）在合适条件下活性酯完全被取代。通过所述方法制备6-氨基己基2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖苷衍生物聚丙烯酰胺凝胶来测试该系统的功效。发现它比先前发表的亲和色谱系统对小麦胚凝集素的纯化更有效，并且以结晶形式获得了小麦胚血凝素。此外，利用pH梯度从亲和凝胶中获得了同工凝集素的部分分离。
