磁分离技术和纳米金比色法用于嗜碱性粒细胞活化试验研究

doi:10.11896/cldb.23030124

材料导报

2024, Vol. 38

Issue (9): 23030124-7 https://doi.org/10.11896/cldb.23030124

高分子与聚合物基复合材料

磁分离技术和纳米金比色法用于嗜碱性粒细胞活化试验研究

王迎迎¹, 刘永欣^2,*, 沈倩², 付婵², 余昌敏^2,*

1 东南大学附属中大医院,南京 210009
2 南京工业大学柔性电子(未来技术)学院,南京 210009

Basophil Activation Test Study Based on Magnetic Separation Technique and Gold Nanoparticle Colorimetry

WANG Yingying¹, LIU Yongxin^2,*, SHEN Qian², FU Chan², YU Changmin^2,*

1 Affiliated Zhongda Hospital,School of Medicine,Southeast University, Nanjing 210009, China
2 School of Flexible Electronics (Future Technologies) (SoFE), Nanjing Tech University, Nanjing 210009, China

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摘要嗜碱性粒细胞活化试验是一种安全、高效的诊断过敏疾病技术,成为临床体外诊断过敏疾病的主要方法之一。目前,嗜碱性粒细胞活化试验主要是利用流式细胞术对细胞进行检测,这种方法通常存在检测时间长、操作繁琐和成本高等缺点。针对这些问题,本研究开发了一种表面修饰HLA-DR抗体的磁珠分离体系,将其与过敏患者的外周血共孵育5 min后,利用磁性分离技术,筛选出表面含有HLA-DR标记物的细胞。测试结果表明,筛选后的嗜碱性粒细胞在全血中的细胞占比从0.2%提升到11.5%,嗜碱性粒细胞活化试验检测灵敏性得到了有效提高。进一步地,本研究构建了CD63适配体修饰的金纳米粒子检测体系,利用CD63适配体特异性识别活化后的嗜碱性粒细胞表面的CD63抗体,使金纳米粒子在嗜碱性粒细胞表面聚集,溶液颜色由红色转变为蓝紫色,实现对激活的嗜碱性粒细胞可视化比色检测,为临床开展嗜碱性粒细胞活化试验提供了新的方法。

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	王迎迎
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	余昌敏

关键词: 比色法金纳米粒子嗜碱性粒细胞激活实验磁分离适配体

Abstract: Basophil activationtest (BAT) is a safe and efficient technique for the diagnosis of allergic diseases, which is one of the main methods for the clinical diagnosis of allergic diseases in vitro. Flow cytometry has been mainly used in BAT. However, this method showed some disadvantages, such as long detection time, cumbersome operation and high cost. To solve these problems, we developed a magnetic bead separation system with surface modified HLA-DR antibody. After co-incubation with peripheral blood of allergic patients for 5 min, cells with HLA-DR markers on the surface were seprated by magnetic separation technology. The results showed that the proportion of basophil cells in whole blood increased from 0.2% to 11.5% after screening, which effectively improved the sensitivity of BAT. Further, we developed a gold nanoparticle-based colorimetry with the CD63 aptamer modification on the surface of gold nanoparticles. Due to the specificity identification of CD63 aptamer with CD63 antibody after activation of basophils, the gold nanoparticles can gather together on the surface of basophils, thus resulting in the color changing from red to violet, which finally realized visual colorimetric detection of the activation of basophils. We herein believe our strategy will provide a new method for clinical BAT.

Key words: colorimetry gold nanoparticles basophil activation test magnetic separation aptamer

出版日期: 2024-05-10 发布日期: 2024-05-13

ZTFLH:

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基金资助: 江苏省高等学校自然科学研究重大项目(19KJA520005);国家卫生健康委核医学重点实验室、江苏省分子核医学重点实验室开放课题(KF202203)

通讯作者: * 刘永欣,2019年6月和2022年6月分别在牡丹江医科大学和南京工业大学获得学士和硕士学位。主要研究方向:生物传感器的制备及在过敏疾病检测中的应用;基于材料学探究海洋超小样品的深部碳循环。发表SCI论文10余篇,包括Talanta、Chin. Chem. Lett.、Anal. Methods等。iamyxliu@njtech.edu.cn
余昌敏,南京工业大学柔性电子(未来技术)学院教授、博士研究生导师。2013年7月,博士毕业于华南理工大学材料学专业。2014年1月至2016年1月在新加坡国立大学开展博士后研究工作。2018年4月加入南京工业大学柔性电子(未来技术)学院。主要研究方向为柔性电子传感器件和有机分子荧光探针的制备及在疾病诊疗中的应用、功能纳米递送体系的构建及生物应用。在Adv. Mater.、Angew. Chem. Int. Ed.、J. Am. Chem. Soc.、Allergy、Small、Chem. Commun.等国际期刊上发表SCI 论文80 余篇。iamcmyu@njtech.edu.cn

作者简介: 王迎迎,2009年东南大学本科毕业。主管护师,现担任东南大学成贤学院内科护理学临床带教;主要研究方向为过敏免疫性疾病干预以及老年慢性病管理等。已发表相关研究论文2篇。

引用本文:

王迎迎, 刘永欣, 沈倩, 付婵, 余昌敏. 磁分离技术和纳米金比色法用于嗜碱性粒细胞活化试验研究[J]. 材料导报, 2024, 38(9): 23030124-7.
WANG Yingying, LIU Yongxin, SHEN Qian, FU Chan, YU Changmin. Basophil Activation Test Study Based on Magnetic Separation Technique and Gold Nanoparticle Colorimetry. Materials Reports, 2024, 38(9): 23030124-7.

链接本文:

http://www.mater-rep.com/CN/10.11896/cldb.23030124 或 http://www.mater-rep.com/CN/Y2024/V38/I9/23030124

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