Magnetic Separation of Antibodies with High Binding Capacity by Site-Directed Immobilization of Protein A-Domains to Bare Iron Oxide Nanoparticles
Magnetic Separation of Antibodies with High Binding Capacity by Site-Directed Immobilization of Protein A-Domains to Bare Iron Oxide Nanoparticles
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DOI：
10.1021/acsanm.1c00487
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发表时间：
2021-05-03
影响因子：
5.9
通讯作者：
Berensmeier, Sonja
中科院分区：
文献类型：
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作者：
Kaveh-Baghbaderani, Yasmin;Allgayer, Raphaela;Berensmeier, Sonja
对纯化抗体的需求不断增长。本研究提出了一种基于纳米粒子的材料，用于有效磁力分离具有高结合能力的免疫球蛋白 G (IgG)。其特点包括： (i) 使用经济高效的裸氧化铁纳米颗粒作为固相，直接固定优化的基于蛋白 A 的配体。不需要额外的化学修饰或活化。 (ii) 使用含有对氧化铁有亲和力的氨基酸的 C 端肽标签促进配体的定向固定。 (iii) 固定配体由蛋白 A 的八个聚合 B 结构域组成。这种用于抗体捕获的亲和吸附剂允许每克颗粒回收高达 418 毫克 IgG，这超过了磁性纳米颗粒和微粒的现有技术水平。配体密度较低的颗粒显示出较高的 IgG 回收率，这使得吸附剂的设计具有成本效益。此外，通过使用兔血清纯化兔多克隆IgG来显示固定配体的选择性。
The demand for purified antibodies is ever-rising. This study presents a nanoparticle-based material for efficient magnetic separation of immunoglobulin G (IgG) with high binding capacity. The characteristics include: (i) Cost-effective bare iron oxide nanoparticles are used as the solid phase on which optimized protein A-based ligands are directly immobilized. An additional chemical modification or activation is not needed. (ii) Oriented immobilization of the ligands is promoted using a C-terminal peptide tag, containing amino acids with an affinity for iron oxide. (iii) The immobilized ligand consists of eight polymerized B-domains of Protein A. This affinity adsorbent for antibody capture allows a recovery of up to 418 mg IgG per gram of particle, which exceeds the state of the art of magnetic nanoparticles as well as microparticles. Particles with a lower ligand density show a higher percentage recovery of IgG, which allows for a cost-effective design of the adsorbent. Furthermore, the selectivity of the immobilized ligand is shown by means of purification of rabbit polyclonal IgG using rabbit serum.