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用于制备IgG结合细菌磁性纳米颗粒的优化方法。

Optimized method for preparation of IgG-binding bacterial magnetic nanoparticles.

作者信息

Grouzdev Denis S, Dziuba Marina V, Kurek Denis V, Ovchinnikov Alexander I, Zhigalova Nadezhda A, Kuznetsov Boris B, Skryabin Konstantin G

机构信息

Faculty of Biology, Moscow State University, Moscow, Russia; Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.

Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.

出版信息

PLoS One. 2014 Oct 15;9(10):e109914. doi: 10.1371/journal.pone.0109914. eCollection 2014.

DOI:10.1371/journal.pone.0109914
PMID:25333971
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4198182/
Abstract

In this study, the optimized method for designing IgG-binding magnetosomes based on integration of IgG-binding fusion proteins into magnetosome membrane in vitro is presented. Fusion proteins Mbb and Mistbb consisting of magnetosome membrane protein MamC and membrane associating protein Mistic from Bacillus subtilis as anchors and BB-domains of Staphylococcus aureus protein A as IgG-binding region were used. With Response Surface Methodology (RSM) the highest level of proteins integration into magnetosome membrane was achieved under the following parameters: pH 8.78, without adding NaCl and 55 s of vortexing for Mbb; pH 9.48, 323 mM NaCl and 55 s of vortexing for Mistbb. Modified magnetosomes with Mbb and Mistbb displayed on their surface demonstrated comparable levels of IgG-binding activity, suggesting that both proteins could be efficiently used as anchor molecules. We also demonstrated that such modified magnetosomes are stable in PBS buffer during at least two weeks. IgG-binding magnetosomes obtained by this approach could serve as a multifunctional platform for displaying various types of antibodies.

摘要

在本研究中，提出了一种基于体外将IgG结合融合蛋白整合到磁小体膜中来设计IgG结合磁小体的优化方法。使用了融合蛋白Mbb和Mistbb，它们由来自枯草芽孢杆菌的磁小体膜蛋白MamC和膜结合蛋白Mistic作为锚定物，以及金黄色葡萄球菌蛋白A的BB结构域作为IgG结合区域。采用响应面法（RSM），在以下参数条件下实现了蛋白整合到磁小体膜中的最高水平：对于Mbb，pH值为8.78，不添加NaCl，涡旋55秒；对于Mistbb，pH值为9.48，323 mM NaCl，涡旋55秒。表面展示有Mbb和Mistbb的修饰磁小体表现出相当水平的IgG结合活性，表明这两种蛋白都可以有效地用作锚定分子。我们还证明，这种修饰的磁小体在PBS缓冲液中至少两周内是稳定的。通过这种方法获得的IgG结合磁小体可以作为展示各种类型抗体的多功能平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/620a8ee3d74f/pone.0109914.g013.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/d6f7ae912e84/pone.0109914.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/691e7be2156f/pone.0109914.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/84bd53cbf4b7/pone.0109914.g009.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/620a8ee3d74f/pone.0109914.g013.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/a119fed68720/pone.0109914.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/562c35eb15db/pone.0109914.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/4198182/fafdbbe57047/pone.0109914.g003.jpg
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