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用于固相肽合成的大孔聚二甲基丙烯酰胺树脂:在Fmoc化学中的应用

Large-pore polydimethylacrylamide resin for solid-phase peptide synthesis: applications in Fmoc chemistry.

作者信息

Sparrow J T, Knieb-Cordonier N G, Obeyseskere N U, McMurray J S

机构信息

Baylor College of Medicine, Houston, TX, USA.

出版信息

Pept Res. 1996 Nov-Dec;9(6):297-304.

PMID:9048423
Abstract

We have synthesized a hydrophilic crosslinked aminoalkyl polydimethylacrylamide-beaded support upon which peptides have been assembled using standard Fmoc chemistry in automated batch-wise equipment. The resin was prepared by the free radical-initiated co-polymerization of N,N-dimethylacryl-amide, N,N'-bisacrylyl-1,3-diaminopropane and a functional monomer N-methacrylyl-1,3-diaminopropane hydrochlorid. After coupling of N-alpha-tert-butyloxycarbonyl-glycine (Boc-glycine), amino acid analyses gave resin loading capacities of 0.66 mmol/g. The resulting polymer was highly swollen by polar solvents including aqueous buffers and had an exclusion limit of 50 kDa for soluble proteins. This resin was found to be an excellent support for peptide synthesis using Fmoc chemistry. Typical purities of crude peptides were 80%-95%, including sequences that failed on conventional polystyrene resins.

摘要

我们合成了一种亲水性交联氨基烷基聚二甲基丙烯酰胺珠状载体,在自动分批设备中使用标准Fmoc化学方法在其上组装了肽。该树脂是通过N,N-二甲基丙烯酰胺、N,N'-双丙烯酰基-1,3-二氨基丙烷和功能性单体N-甲基丙烯酰基-1,3-二氨基丙烷盐酸盐的自由基引发共聚合制备的。在偶联N-α-叔丁氧羰基甘氨酸(Boc-甘氨酸)后,氨基酸分析给出的树脂负载量为0.66 mmol/g。所得聚合物在包括水性缓冲液在内的极性溶剂中高度溶胀,对可溶性蛋白质的排阻极限为50 kDa。发现该树脂是使用Fmoc化学方法进行肽合成的优良载体。粗肽的典型纯度为80%-95%,包括在传统聚苯乙烯树脂上合成失败的序列。

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