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使用 κ-卡拉胶修饰的杂化磁性纳米粒子从兔血清中回收免疫球蛋白 G。

Recovery of immunoglobulin G from rabbit serum using κ-carrageenan-modified hybrid magnetic nanoparticles.

机构信息

CICECO - Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal.

CICECO - Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal.

出版信息

Int J Biol Macromol. 2020 May 1;150:914-921. doi: 10.1016/j.ijbiomac.2020.02.135. Epub 2020 Feb 14.

Abstract

Immunoglobulin G (IgG) has been used in the treatment of cancer, autoimmune diseases and neurological disorders, however, the current technologies to purify and recover IgG from biological media are of high-cost and time-consuming, resulting in high-cost products. In this sense, the search for cost-effective technologies to obtain highly pure and active IgG is highly required. The present work proposes a simple and efficient method for the purification and recovery of IgG from rabbit serum using magnetic iron oxide nanoparticles (magnetite, FeO) coated with hybrid shells of a siliceous material modified with the anionic polysaccharide κ-carrageenan. Experimental parameters such as pH, contact time between the hybrid magnetic nanoparticles (HMNPs) and rabbit serum, and total protein concentration or dilution factor of serum were evaluated. The best results were achieved at pH 5.0, with a contact time of 60 min and using a rabbit serum with a total protein concentration of 4.8 mg·mL. Under these conditions, it was obtained an IgG purification factor and adsorption yield onto the HMNPs of 3.0 and 90%, respectively. The desorption of IgG from the HMNPs was evaluated using two strategies: a KCl aqueous solution and buffered aqueous solutions. Comparing to the initial rabbit serum, an IgG purification factor of 2.7 with a recovery yield of 74% were obtained using a buffered aqueous solution at pH 7.0. After desorption, the secondary structure of IgG and other proteins was evaluated by circular dichroism and no changes in the secondary structure were observed, meaning that the IgG integrity is kept after the adsorption and desorption steps. In summary, the application of HMNPs in the purification of IgG from serum samples has a high potential as a new downstream platform.

摘要

免疫球蛋白 G(IgG)已被用于癌症、自身免疫性疾病和神经紊乱的治疗,但目前从生物介质中纯化和回收 IgG 的技术成本高、耗时,导致产品成本高。从这个意义上说,迫切需要寻找经济高效的技术来获得高纯度和高活性的 IgG。本工作提出了一种从兔血清中纯化和回收 IgG 的简单高效方法,使用的是经过阴离子多糖 κ-卡拉胶修饰的硅质材料的混合壳涂覆的磁性氧化铁纳米粒子(磁铁矿,FeO)。实验参数如 pH 值、混合磁性纳米粒子(HMNPs)与兔血清之间的接触时间以及血清中的总蛋白浓度或稀释因子都进行了评估。最佳结果在 pH 值为 5.0 时获得,接触时间为 60 分钟,使用的是总蛋白浓度为 4.8mg·mL 的兔血清。在这些条件下,获得了 IgG 纯化因子和吸附到 HMNPs 的产率分别为 3.0 和 90%。通过两种策略评估了 IgG 从 HMNPs 上的解吸:KCl 水溶液和缓冲水溶液。与初始兔血清相比,使用 pH 值为 7.0 的缓冲水溶液获得了 IgG 纯化因子 2.7 和回收率 74%。解吸后,通过圆二色性评估了 IgG 和其他蛋白质的二级结构,未观察到二级结构发生变化,这意味着在吸附和解吸步骤后 IgG 的完整性得以保持。总之,HMNPs 在从血清样品中纯化 IgG 方面的应用具有作为新的下游平台的高潜力。

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