Penke B, Zarándi M, Varga J R, Tóth G K, Kovács K, Szajáni B
J Chromatogr. 1986 Apr 11;376:307-14. doi: 10.1016/s0378-4347(00)80847-6.
A new strategy was devised for the targeted immobilization of ligands on aminohexyl- and carboxyhexyl-agarose. Selectively protected neurotransmitter amino acids and neuropeptides were coupled to amino or carboxyl group-containing agarose derivatives using activated esters, mixed anhydrides or carbodiimides. After coupling, agarose beads were dehydrated and the protecting groups were cleaved in non-aqueous media with acids (trifluoroacetic acid, formic acid). Agarose beads were rehydrated and applied for affinity chromatography and cell surface recognition. The same compounds were coupled to derivatized polyacrylamide beads containing primary amino (Acrylex A), acyl hydrazide (Acrylex AH-100) or carboxyl (Acrylex C-100) groups. Protecting groups were removed by acidolytic cleavage. Oxytocin, vasopressin, tetra- and pentagastrin, cholecystokinin, leucine-enkephalin and carboxyl-bearing derivatives of the neurotransmitters noradrenaline, dopamine, histamine, serotonin, acetylcholine and gamma-aminobutyric acid were immunobilized on agarose and on derivatized polyacrylamide gels.
设计了一种新策略，用于将配体靶向固定在氨基己基和羧基己基琼脂糖上。使用活性酯、混合酸酐或碳二亚胺，将选择性保护的神经递质氨基酸和神经肽与含氨基或羧基的琼脂糖衍生物偶联。偶联后，将琼脂糖珠脱水，并在非水介质中用酸（三氟乙酸、甲酸）裂解保护基团。使琼脂糖珠重新水化，并用于亲和色谱和细胞表面识别。将相同的化合物偶联到含有伯氨基（Acrylex A）、酰肼（Acrylex AH - 100）或羧基（Acrylex C - 100）基团的衍生化聚丙烯酰胺珠上。通过酸解裂解去除保护基团。将催产素、加压素、四肽和五肽胃泌素、胆囊收缩素、亮氨酸脑啡肽以及神经递质去甲肾上腺素、多巴胺、组胺、5 - 羟色胺、乙酰胆碱和γ - 氨基丁酸的含羧基衍生物免疫固定在琼脂糖和衍生化聚丙烯酰胺凝胶上。
