Homodimeric Protein–Polymer Conjugates via the Tetrazine–trans-Cyclooctene Ligation

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• 作者：Maltish M. Lorenzo ; Caitlin G. Decker ; Muhammet U. Kahveci ; Samantha J. Paluck ; Heather D. Maynard
• 刊名：Macromolecules
• 出版年：2016
• 出版时间：January 12, 2016
• 年：2016
• 卷：49
• 期：1
• 页码：30-37
• 全文大小：442K
• ISSN：1520-5835

文摘

Tetrazine end-functionalized telechelic polymers were synthesized by controlled radical polymerization (CRP) and employed to generate T4 lysozyme homodimers. Mutant T4 lysozyme (V131C), containing a single surface-exposed cysteine, was modified with a protein-reactive trans-cyclooctene (T4L-TCO). Reversible addition–fragmentation chain transfer (RAFT) polymerization yielded poly(N-isopropylacrylamide) (pNIPAAm) with a number-average molecular weight (M_n by ¹H NMR) of 2.0 kDa and a dispersity (? by GPC) of 1.05. pNIPAAm was then modified at both ends by postpolymerization with 6-methyltetrazine. For comparison, 2.0 kDa bis-tetrazine poly(ethylene glycol) (PEG) and 2.0 kDa bis-maleimide pNIPAAm were synthesized. Ligation of T4L-TCO to bis-tetrazine pNIPAAm or bis-tetrazine PEG resulted in protein homodimer in 38% yield and 37% yield, respectively, after only 1 h, whereas bis-maleimide pNIPAAm resulted in only 5% yield of dimer after 24 h. This work illustrates the advantage of employing tetrazine ligation over maleimide thiol–ene chemistry for the synthesis of protein homodimer conjugates.