Hybrid alginate–protein cryogel beads: efficient and sustainable bio-based materials to purify immunoglobulin G antibodies,Green Chemistry

当前位置： X-MOL 学术 › Green Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Hybrid alginate–protein cryogel beads: efficient and sustainable bio-based materials to purify immunoglobulin G antibodies
Green Chemistry ( IF 9.3 ) Pub Date : 2020-02-26 , DOI: 10.1039/c9gc04449c
Mukesh Sharma _{1,

2,

3,

4,

5} , Ana Paula M. Tavares _{1,

2,

3,

4,

5} , João C. F. Nunes _{1,

2,

3,

4,

5} , Nripat Singh _{6,

7,

8} , Dibyendu Mondal _{8,

9,

10,

11} , Márcia C. Neves _{1,

2,

3,

4,

5} , Kamalesh Prasad _{6,

7,

8} , Mara G. Freire _{1,

2,

3,

4,

5}

Affiliation

Antibodies present in mammal's serum are of high relevance for therapeutic purposes, particularly in passive immunization and in the treatment of some chronic diseases. However, their widespread use is still compromised by the requirement of several process steps for their purification and the difficulty in keeping antibodies stable to guarantee their therapeutic efficiency. These challenges significantly contribute to the current high cost of biopharmaceuticals, namely antibodies such as immunoglobulin G (IgG). Accordingly, the development of effective and sustainable purification strategies for antibodies and other biopharmaceuticals is in critical demand to decrease economic, environmental and health burdens. Herein, bio-based and low-cost hybrid alginate–protein cryogel beads were prepared, characterized, and applied as novel adsorbent materials for the purification of IgG from human serum. It is shown that hybrid materials are more efficient than the respective alginate beads since the presence of proteins increases the material selectivity for IgG, which is due to the specific interactions occurring between the target antibody and amino acid residues in the hybrid materials. Several operating conditions, such as pH, adsorption time and serum concentration, were optimized to improve the recovery yield and purity of IgG. Adsorption isotherms were determined to infer the adsorption mechanism of IgG onto the cryogel beads and to determine their adsorption capacity (175 mg of IgG per g of cryogel beads). Under the optimized conditions, IgG can be recovered from the hybrid materials using buffered aqueous solutions, with a purity of 80% and a recovery yield of 91%. The stability and integrity of the antibody are retained after the desorption step. Finally, the regeneration and reuse of the cryogel beads were evaluated, with no losses in the IgG adsorption performance and antibody stability. Although significant efforts have been put on the development of novel affinity ligands to replace the standard chromatographic methods to purify IgG, this work demonstrates the potential of bio-based and low-cost hybrid materials as promising alternatives, in which proteins can be used to improve the material selectivity.

中文翻译：

藻酸盐-蛋白质混合冻凝胶珠子：高效且可持续的生物基材料，可纯化免疫球蛋白G抗体

哺乳动物血清中存在的抗体对于治疗目的具有高度相关性，特别是在被动免疫和某些慢性疾病的治疗中。然而，它们的广泛使用仍然受到其纯化的几个处理步骤的要求以及难以保持抗体稳定以确保其治疗效率的困难。这些挑战极大地导致了目前生物药物的高成本，即抗体，例如免疫球蛋白G（IgG）。因此，迫切需要开发有效和可持续的抗体和其他生物药物纯化策略，以减少经济，环境和健康负担。在此，制备了基于生物的低成本低成本藻酸盐-蛋白质杂交凝胶珠，并用作从人血清中纯化IgG的新型吸附材料。结果表明，杂化材料比相应的藻酸盐珠更有效，因为蛋白质的存在增加了对IgG的材料选择性，这是由于目标抗体与杂化材料中的氨基酸残基之间发生了特定的相互作用。优化了几个操作条件，例如pH，吸附时间和血清浓度，以提高IgG的回收率和纯度。测定吸附等温线以推断IgG在冷冻凝胶珠上的吸附机理并确定其吸附能力（每克冷冻凝胶珠为175mg IgG）。在最佳条件下，可以使用缓冲水溶液从杂种材料中回收IgG，纯度为80％，回收率为91％。解吸步骤后保留抗体的稳定性和完整性。最后，评估了冷冻凝胶珠的再生和再利用，而IgG吸附性能和抗体稳定性没有损失。尽管已经在开发新型亲和配体方面做出了巨大努力，以取代用于纯化IgG的标准色谱方法，但这项工作证明了基于生物和低成本的杂化材料作为有前途的替代品的潜力，其中可以使用蛋白质来改善蛋白质材料的选择性。

更新日期：2020-02-26

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11