Hearn M T, Harris E L, Bethell G S, Hancock W S, Ayers J A
J Chromatogr. 1981 Nov 20;218:509-18. doi: 10.1016/s0021-9673(00)82076-2.
The characteristics of ligands immobilised into 1,1'-carbonyldiimidazole-treated cross-linked agarose have been further evaluated. Since the intermediate activated matrix (an imidazolyl carbamate-agarose) is susceptible to nucleophilic attack by free amino groups, but is relatively stable to oxygen nucleophiles, ligands ranging from simple organic primary amines, amino acids, proteins and other biological substances, which contain amino group functionality, can be bound to the agarose via a N-alkylcarbamate (urethane) linkage. This covalent linkage has been found to exhibit good chemical stability to mildly acidic and basic elution conditions. The use of 1,1'-carbonyldiimidazole-activated agarose in the biospecific affinity chromatography of immunoglobulins, present in normal and pathological sera, is described.
对固定在1,1'-羰基二咪唑处理的交联琼脂糖中的配体特性进行了进一步评估。由于中间活化基质(咪唑基氨基甲酸酯-琼脂糖)易受游离氨基的亲核攻击,但对氧亲核试剂相对稳定,因此含有氨基官能团的简单有机伯胺、氨基酸、蛋白质和其他生物物质等配体,可以通过N-烷基氨基甲酸酯(脲)键与琼脂糖结合。已发现这种共价键在温和的酸性和碱性洗脱条件下具有良好的化学稳定性。本文描述了1,1'-羰基二咪唑活化琼脂糖在正常和病理血清中免疫球蛋白的生物特异性亲和色谱中的应用。