A versatile method for the conjugation of proteins and peptides to poly[2-(dimethylamino)ethyl methacrylate].
A versatile method for the conjugation of proteins and peptides to poly[2-(dimethylamino)ethyl methacrylate].
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一种将蛋白质和肽缀合至聚[2-(二甲基氨基)乙基甲基丙烯酸酯]的通用方法。
DOI：
10.1021/bc980126
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发表时间：
1999
影响因子：
4.7
通讯作者：
W. Hennink
中科院分区：
文献类型：
--
作者：
W. N. van Dijk;P. van de Wetering;W. Hinrichs;L. Hofmeyer;R. Liskamp;D. Crommelin;W. Hennink
采用自由基聚合法合成了甲基丙烯酸二甲氨基乙酯（DMAEMA）与甲基丙烯酸氨基乙酯（AEMA）的无规共聚物。引入的伯氨基的量可以通过AEMA与DMAEMA的进料比来控制，并且在2至6摩尔%之间变化。随后，在用N-琥珀酰亚胺基3-（2-吡啶基二硫代）丙酸酯（SPDP）衍生化步骤中引入受保护的巯基，随后用二硫苏糖醇（DTT）处理。将获得的硫醇化p（DMAEMA-共-AEMA）通过二硫键与转铁蛋白（Tf）或mAb 323/A3的F（ab '）片段缀合。此外，核定位信号（NLS）十肽Gly-Pro-Lys-Lys-Lys-Arg-Lys-Val-Glu-Asp-NH（2）的马来酰亚胺衍生物通过硫醚键与硫醇化聚合物偶联。通过GPC（Tf）、SDS-PAGE [F（ab '）]或（1）H NMR（NLS肽）测定，Tf缀合物的偶联效率为90-95%，F（ab'）缀合物和NLS缀合物的偶联效率超过95%。本文所述的合成策略是制备蛋白质和肽与pDMAEMA特别是缀合物的通用方法。该方法一般可用于合成蛋白质-聚甲基丙烯酸酯缀合物。
Random copolymers of 2-(dimethylamino) ethyl methacrylate (DMAEMA) with aminoethyl methacrylate (AEMA) were synthesized by radical polymerization. The amount of incorporated primary amino groups could be controlled by the feed ratio of AEMA to DMAEMA, and was varied from 2 to 6 mol %. Subsequently, protected thiol groups were introduced in a derivatization step with N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) and subsequent treatment with dithiothreitol (DTT). The obtained thiolated p(DMAEMA-co-AEMA) was conjugated to transferrin (Tf) or the F(ab') fragment of mAb 323/A3 via a disulfide linkage. Moreover, the maleimide derivative of the nuclear localization signal (NLS) decapeptide Gly-Pro-Lys-Lys-Lys-Arg-Lys-Val-Glu-Asp-NH(2) was coupled to the thiolated polymer via a thioether linkage. The coupling efficiency, as determined by GPC (Tf), SDS-PAGE [F(ab')], or (1)H NMR (NLS peptide) was 90-95% for the Tf conjugate, and more than 95% for the F(ab') conjugate and the NLS conjugate. The synthetic strategy described in this paper is a universal method for the preparation of conjugates of proteins and peptides with pDMAEMA in particular. This method can possibly be used to synthesize protein-polymethacrylate conjugates in general.