Immunomagnetic‐bead enriched culturomics (IMBEC) for isolating pathobionts from feces of colorectal cancer patients
Immunomagnetic‐bead enriched culturomics (IMBEC) for isolating pathobionts from feces of colorectal cancer patients
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DOI：
10.1002/imt2.100
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发表时间：
2023-04
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通讯作者：
Zi-Yao Huang;Yuxiao Chang;Kun Hao;Ya-fang Tan;Lei Ding;Likun Wang;Zhen Wang;Zhiyuan Pan;Hong Gao;Jiahong Wu;Yubing Zhu;Q. Gao;Y. Bi;Ruifu Yang
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作者：
Zi-Yao Huang;Yuxiao Chang;Kun Hao;Ya-fang Tan;Lei Ding;Likun Wang;Zhen Wang;Zhiyuan Pan;Hong Gao;Jiahong Wu;Yubing Zhu;Q. Gao;Y. Bi;Ruifu Yang
摘要培养组学利用不同的培养条件获得不同类型的细菌和新种。然而，目前的培养组学缺乏一种高效的方法来分离特定的病原体。免疫磁珠技术是利用与抗体结合的磁珠捕获抗原来实现靶细胞富集的一种替代方法。在这项研究中，我们开发了一种新的方法——免疫磁珠富集培养组学（IMBEC），该方法从结直肠癌（CRC）患者的粪便样本中纯化磁珠偶联抗体，用于富集和分离潜在的病原体。通过优化偶联剂、NaCl和洗涤剂的浓度，开发了一种通过免疫磁捕获富集潜在病原体的方案。比较了抗体包被磁珠（抗体组）和未偶联的空白磁珠（空白组）对病原菌富集的效果。为了确定两组分离物的促炎潜能，我们研究了它们诱导THP‐1巨噬细胞产生细胞因子的能力。采用该方案从10例结直肠癌患者的粪便样本中分离细菌，同时与空白组分离的细菌进行比较。两组共分离细菌209种，其中抗体组173种，空白组160种，两组124种。从抗体组分离的细菌比从空白组分离的细菌产生更多的促炎细胞因子。IMBEC是一种很有前途的方法，可以相对特异性地分离出潜在的病原体。
Abstract Culturomics employs various cultivating conditions to obtain different types of bacteria and new species. However, current culturomics lacks a highly efficient method for isolating specific pathobionts. Immunomagnetic bead technology, which uses magnetic beads conjugated with antibodies for capturing the antigen to realize enrichment of the targets, has been employed as an alternative method. In this study, we developed a novel method, immunomagnetic bead‐enriched culturomics (IMBEC), in which magnetic bead‐conjugated antibodies purified from the fecal samples of patients with colorectal cancer (CRC) were used to enrich and isolate potential pathobionts. A protocol for enriching potential pathobionts via immunomagnetic capture was developed by optimizing the concentrations of coupling reagents, NaCl, and detergent. The efficacy of pathobiont enrichment was compared between antibody‐coated magnetic beads (antibody group) and nonconjugated blank magnetic beads (blank group). To determine the proinflammatory potential of isolates from both groups, we investigated their ability to induce cytokine production in THP‐1 macrophages. This protocol was employed for isolating bacteria from 10 fecal samples of patients with CRC, which were simultaneously compared with those isolated from the blank group. A total of 209 bacterial species were isolated from both groups, including 173 from the antibody group, 160 from the blank group, and 124 from both groups. Bacteria isolated from the antibody group produced more proinflammatory cytokines than those isolated from the blank group. IMBEC is a promising method for relatively specific isolation of potential pathobionts for a particular disease of interest.