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高性能金黄色葡萄球菌蛋白A亲和层析介质的制备、性能与应用
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江南大学粮食发酵与食品生物制造国家工程研究中心，江苏 无锡 214122

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夏海锋（1979—），男，博士，副教授，硕士研究生导师，主要从事发酵工程、生化工程、生物分离工程以及学科交叉研究。E-mail：hfxia@jiangnan.edu.cn

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Preparation, Performance, and Application of a High-performance Affinity Chromatography Medium Based on Staphylococcus aureus Protein A
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National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China

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    摘要:

    目的 金黄色葡萄球菌蛋白A(Staphylococcus aureus protein A，SPA）对免疫球蛋白G（immunoglobulin G，IgG）具有高度的特异性，已广泛应用于单克隆抗体的纯化。然而，商品介质价格高昂，限制了其进一步发展。为解决此问题，该研究试图获得高性能的国产SPA亲和层析介质，以替代进口层析介质。方法 对重组菌诱导表达获得的SPA，经Ni2+纯化后偶联至Rigose HF基质，以获得自制亲和层析介质。结果 自制亲和层析介质静态饱和吸附量为115.5 mg/mL（以介质体积计），动态吸附量为58.6 mg/mL（以介质体积计）；利用自制亲和层析介质分离纯化单克隆抗体样品，纯度在95%以上，回收率不低于90%，耐碱性、重复使用性能以及纯化单克隆抗体样品的效果等同或优于商品介质。结论 该研究成功构建了SPA重组菌，通过优化偶联反应条件，确定了自制亲和层析介质的最佳制备条件，具有一定的研究意义和商业应用价值。

    Abstract:

    Objective Staphylococcus aureus protein A (SPA) is widely used for the purification of monoclonal antibodies due to its high specificity for immunoglobulin G (IgG). However, the high costs of commercial media have limited the further development of SPA. To address this issue, this paper aims to prepare a high-performance affinity chromatography medium based on SPA as the substitute of introduced affinity chromatography media.Method SPA expressed by the recombinant strain was purified via Ni2+ affinity chromatography and then coupled to the Rigose HF matrix to prepare an self-prepared affinity chromatography medium.Result The self-prepared medium had a static saturated adsorption capacity of 115.5 mg/mL (based on the volume of the medium) and a dynamic adsorption capacity of 58.6 mg/mL (based on the volume of the medium). When being used for the separation and purification of monoclonal antibody samples, the self-prepared affinity chromatography medium achieved the purity of over 95%, with a recovery rate of no less than 90%. The medium demonstrated comparable or superior alkali tolerance, reusability, and purification performance for monoclonal antibody samples to commercial media.Conclusion A recombinant strain expressing SPA is successfully constructed, and the optimal preparation conditions of the self-prepared affinity chromatography medium are determined by optimization of the coupling conditions. The self-prepared affinity chromatography medium demonstrates research significance and commercial application value to a certain extent.

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郭子豪,闫朝清,夏海锋.高性能金黄色葡萄球菌蛋白A亲和层析介质的制备、性能与应用[J].食品与生物技术学报,2025,(5):137-145.

GUO Zihao, YAN Chaoqing, XIA Haifeng. Preparation, Performance, and Application of a High-performance Affinity Chromatography Medium Based on Staphylococcus aureus Protein A[J]. Journal of Food Science and Biotechnology,2025,(5):137-145.

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  • 收稿日期:2022-12-20
  • 最后修改日期:2023-03-16
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  • 在线发布日期: 2025-09-02
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