TEMPO氧化的纳米原纤化纤维素作为生物活性分子的高密度载体,Biomacromolecules

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TEMPO氧化的纳米原纤化纤维素作为生物活性分子的高密度载体
Biomacromolecules ( IF 5.5 ) Pub Date : 2015-10-13 00:00:00 , DOI: 10.1021/acs.biomac.5b01100
Ramon Weishaupt ₁ , Gilberto Siqueira ₂ , Mark Schubert ₂ , Philippe Tingaut ₂ , Katharina Maniura-Weber ₁ , Tanja Zimmermann ₂ , Linda Thöny-Meyer ₁ , Greta Faccio ₁ , Julian Ihssen ₁

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特定生物分子的受控和有效固定是一项关键技术，可将新的有利功能引入适合生物医学应用的材料。在这里，我们描述了一种创新且高效的两步方法，用于将各种生物分子（包括小肽和酶）稳定固定在TEMPO氧化的纳米原纤化纤维素（TO-NFC）上。通过TEMPO氧化将羧酸酯基团引入NFC中提供了高表面密度的负电荷，该负电荷能够驱动生物分子的吸附并参与与1-乙基-3- [3-（二甲基氨基）丙基]碳二亚胺的共价交联反应（EDAC）和戊二醛（Ga）化学。每毫克TO-NFC最多可通过静电相互作用将0.27μmol不同的生物分子固定化。额外的化学交联步骤可防止80％以上的这些分子解吸。使用半胱氨酸蛋白酶木瓜蛋白酶作为模型，获得了高活性的木瓜蛋白酶-TO-NFC偶联物。木瓜蛋白酶固定后，保留了40％的初始酶活性，并增加了对于共价固定化酶，k _cat从213到> 700 s ^–1。通过使定义明确的具有高功能密度的混合生物材料成为可能，这项工作中介绍的方法论扩展了TO-NFC在生物医学领域的应用范围。

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TEMPO-Oxidized Nanofibrillated Cellulose as a High Density Carrier for Bioactive Molecules

Controlled and efficient immobilization of specific biomolecules is a key technology to introduce new, favorable functions to materials suitable for biomedical applications. Here, we describe an innovative and efficient, two-step methodology for the stable immobilization of various biomolecules, including small peptides and enzymes onto TEMPO oxidized nanofibrillated cellulose (TO-NFC). The introduction of carboxylate groups to NFC by TEMPO oxidation provided a high surface density of negative charges able to drive the adsorption of biomolecules and take part in covalent cross-linking reactions with 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDAC) and glutaraldehyde (Ga) chemistry. Up to 0.27 μmol of different biomolecules per mg of TO-NFC could be reversibly immobilized by electrostatic interaction. An additional chemical cross-linking step prevented desorption of more than 80% of these molecules. Using the cysteine-protease papain as model, a highly active papain-TO-NFC conjugate was achieved. Once papain was immobilized, 40% of the initial enzymatic activity was retained, with an increase in k_cat from 213 to >700 s^–1 for the covalently immobilized enzymes. The methodology presented in this work expands the range of application for TO-NFC in the biomedical field by enabling well-defined hybrid biomaterials with a high density of functionalization.

更新日期：2015-10-13

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