Water-soluble carrier proteins having carboxyl spacer groups.
作者:MIKIO OHNISHI、HIROYUKI SUGIMOTO、HIDENORI YAMADA、TAIJI IMOTO、KIYOSHI ZAITSU、YOSUKE OHKURA
DOI:10.1248/cpb.33.2871
日期:——
Three water-soluble bovine serum albumin derivatives with carboxyl spacer groups have been synthesized as carrier proteins for direct preparation of conjugates between proteins and low molecular haptens with amino groups. N-Glycylaminomethanesulfonic acid, a newly developed solubilizing reagent for proteins, was first introduced into bovine serum albumin through its carboxyl groups. After the methylation of carboxyl groups remaining unreacted in the modified albumin, N-succinylglycine and N-bromoacetyl-β-alanine, both carboxyl spacer reagents, were reacted with the amino or imidazole groups of the modified albumin, respectively, to give two water-soluble albumin derivatives. Another water-soluble albumin derivatives. Another water-soluble albumin derivative was obtained by treating the modified albumin with N-bromoacetyl-β-alanine followed by amidination of the remaining amino groups of the albumin with methylacetimidate. Glycine ethyl ester, selected as a model hapten that has an amino group, was reacted with these albumin derivatives to form conjugates, and 4-15 ester moieties can be incorporated into one molecule of the albumin derivatives without loss of their water-solubility.
合成了三种带有羧基间隔基团的水溶性牛血清白蛋白衍生物作为载体蛋白,用于直接制备蛋白质与带有氨基的低分子半抗原之间的缀合物。 N-甘氨酰氨基甲磺酸是一种新开发的蛋白质溶解试剂,首次通过其羧基引入牛血清白蛋白中。修饰白蛋白中未反应的羧基甲基化后,N-琥珀酰甘氨酸和N-溴乙酰基-β-丙氨酸(两种羧基间隔试剂)分别与修饰白蛋白的氨基或咪唑基团反应,得到两种水-可溶性白蛋白衍生物。另一种水溶性白蛋白衍生物。另一种水溶性白蛋白衍生物是通过用N-溴乙酰基-β-丙氨酸处理修饰的白蛋白,然后用乙酰亚氨酸甲酯对白蛋白的剩余氨基进行脒化而获得的。选择甘氨酸乙酯作为具有氨基的模型半抗原,与这些白蛋白衍生物反应形成缀合物,并且4-15个酯部分可以掺入到白蛋白衍生物的一个分子中而不损失其水溶性。