Tag-Free Site-Specific BMP-2 Immobilization with Long-Acting Bioactivities via a Simple Sugar–Lectin Interaction,ACS Biomaterials Science & Engineering

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Tag-Free Site-Specific BMP-2 Immobilization with Long-Acting Bioactivities via a Simple Sugar–Lectin Interaction
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-02-26 , DOI: 10.1021/acsbiomaterials.9b01730
Honglei Wang , Xue Qu , Zheng Zhang ₁ , Miao Lei , Haoqi Tan , Chunyan Bao , Shaoliang Lin , Linyong Zhu , Joachim Kohn ₁ , Changsheng Liu

Affiliation

The construction of a biomaterial matrix with biological properties is of great importance to developing functional materials for clinical use. However, the site-specific immobilization of growth factors to endow materials with bioactivities has been a challenge to date. Considering the wide existence of glycosylation in mammalian proteins or recombinant proteins, we establish a bioaffinity-based protein immobilization strategy (bioanchoring method) utilizing the native sugar–lectin interaction between concanavalin A (Con A) and the oligosaccharide chain on glycosylated bone morphogenetic protein-2 (GBMP-2). The interaction realizes the site-specific immobilization of GBMP-2 to a substrate modified with Con A while preserving its bioactivity in a sustained and highly efficient way, as evidenced by its enhanced ability to induce osteodifferentiation compared with that of the soluble GBMP-2. Moreover, the surface with Con A-bioanchored GBMP-2 can be reused to stimulate multiple batches of C2C12 cells to differentiate almost to the same degree. Even after 4 month storage at 4 °C in phosphate-buffered saline (PBS), the Con A-bioanchored GBMP-2 still maintains the bioactivity to stimulate the differentiation of C2C12 cells. Furthermore, the ectopic ossification test proves the in vivo bioactivity of bioanchored GBMP-2. Overall, our results demonstrate that the tag-free and site (i.e., sugar chain)-specific protein immobilization strategy represents a simple and generic alternative, which is promising to apply for other glycoprotein immobilization and application. It should be noted that although the lectin we utilized can only bind to d-mannose/d-glucose, the diversity of the lectin family assures that a specific lectin could be offered for other sugar types, thus expanding the applicable scope further.

中文翻译：

通过简单的糖-凝集素相互作用，具有长效生物活性的无标签特定位点BMP-2固定化

具有生物学特性的生物材料基质的构建对于开发用于临床的功能材料非常重要。然而，迄今为止，生长因子的位点固定化赋予材料具有生物活性的挑战。考虑到哺乳动物蛋白或重组蛋白中糖基化的广泛存在，我们利用伴刀豆球蛋白A（Con A）和糖基化骨形态发生蛋白-上的寡糖链之间的天然糖-凝集素相互作用，建立了基于生物亲和力的蛋白质固定策略（生物固定方法） 2（GBMP-2）。相互作用可以实现GBMP-2的位点特异性固定在Con A修饰的底物上，同时以持续高效的方式保留其生物活性，与可溶性GBMP-2相比，其增强的诱导骨分化能力证明了这一点。而且，具有Con A-生物锚定GBMP-2的表面可以重复使用，以刺激多批C2C12细胞，使其分化程度几乎相同。即使在磷酸盐缓冲盐水（PBS）中于4°C储存4个月后，Con A-生物锚定的GBMP-2仍保持刺激C2C12细胞分化的生物活性。此外，异位骨化测试证明了生物锚定GBMP-2的体内生物活性。总的来说，我们的结果表明，无标签和位点（即糖链）特异性蛋白固定策略代表了一种简单而通用的替代方法，有望应用于其他糖蛋白的固定和应用。d-甘露糖/ d-葡萄糖，凝集素家族的多样性确保可以为其他糖类型提供特定的凝集素，从而进一步扩大了适用范围。

更新日期：2020-02-26

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