Protein release from biodegradable polyHPMA-lysozyme conjugates resulting in bioactivity enhancement.
Protein release from biodegradable polyHPMA-lysozyme conjugates resulting in bioactivity enhancement.
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DOI：
10.1002/asia.201000729
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发表时间：
2011-06
期刊：
影响因子：
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通讯作者：
Lei Tao;Gaojian Chen;Li-Xiang Zhao;Jiangtao Xu;E. Huang;Aiping Liu;C. Marquis;T. P. Davis
中科院分区：
文献类型：
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作者：
Lei Tao;Gaojian Chen;Li-Xiang Zhao;Jiangtao Xu;E. Huang;Aiping Liu;C. Marquis;T. P. Davis
合成了一种新型的可生物降解的噻唑烷-2-酮功能链转移剂，并将其作为可逆的额外断裂链转移剂，制备了具有预定分子量和窄多分散性的半遥爪聚N-（2-羟丙基）甲基丙烯酰胺（polyHPMAs）。蛋白质反应基团噻唑烷-2-基位于通过可生物降解的二硫键固定的聚合物链末端。随后，通过利用噻唑烷-2-酮官能团与蛋白质表面上的胺残基之间的反应形成共价酰胺键，将功能性聚HPMA链缀合至蛋白质（溶菌酶）。以溶壁微球菌细胞为底物，研究了溶菌酶-聚HPMA偶联物的体外生物活性。溶菌酶的生物活性显着降低后的缀合程序。然而，从生物缀合物中裂解聚合物链（在还原条件下）产生游离蛋白质和生物活性的显著恢复。通过皮下注射到小鼠中进行体内测试，并且清楚地证明当与天然蛋白质相比时，蛋白质-聚合物缀合物的蛋白水解降解降低，表明通过缀合策略的有效蛋白质保护。这种生物可逆的缀合方法允许在蛋白质保护和有效的生物活性维持之间取得平衡。
A novel biodegradable thiazolidine-2-thione functional chain transfer agent was synthesized and employed as a reversible additional fragmentation chain transfer agent to prepare well-defined semitelechelic poly-N-(2-hydroxypropyl) methacrylamides (polyHPMAs) with predetermined molecular weights and narrow polydispersities. The protein reactive group, thiazolidine-2-thione, was located at the polymer chain ends fixed by biodegradable disulfide bonds. The functional polyHPMA chains were subsequently conjugated to protein (lysozyme) by exploiting reactions between the thiazolidine-2-thione functionality and amine residues on the protein surface to form covalent amide linkages. The in vitro bioactivities of the lysozyme-polyHPMA conjugates were assessed by using Micrococcus lysodeikticus cells as substrates. The lysozyme bioactivity was significantly reduced following the conjugation procedure. However, cleavage of the polymer chains from the bioconjugates (under reducing conditions) yielded free protein and a remarkable recovery of bioactivity. In vivo tests were performed by subcutaneous injection into mice and clearly demonstrated decreased proteolytic degradation for the protein-polymer conjugate when compared with native protein, indicating effective protein protection through a conjugation strategy. This bioreversible approach to conjugation allows for a balance to be made between protein protection and effective bioactivity maintenance.