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注射器内实验室自动化微型磁珠与抗SARS-CoV2抗体生物共轭反应

Lab-in-Syringe Automated Miniaturized Bioconjugation of Magnetic Beads with Anti-SARS-CoV2 Antibodies.

作者信息

Svobodova Zuzana, Krizova Lucie, Matejkova Nikola, Smela Denisa, Beranek Martin, Bilkova Zuzana, Horstkotte Burkhard

机构信息

Charles University, Faculty of Pharmacy, Department of Biological and Medical Sciences, Hradec Kralove 500 09, Czech Republic.

Charles University, Faculty of Pharmacy, Department of Analytical Chemistry, Hradec Kralove 500 03, Czech Republic.

出版信息

ACS Omega. 2025 Jul 21;10(29):31359-31367. doi: 10.1021/acsomega.4c11126. eCollection 2025 Jul 29.

Abstract

We present the first automated synthesis of magnetic immunosorbents (MIS) using a lab-in-syringe (LIS) platform, facilitating antibody bioconjugation to magnetic beads via carbodiimide-mediated covalent binding. This approach is an efficient, reproducible alternative to traditional manual methods, minimizing pipetting steps, vortexing, and incubation with a reduced handling bias. Utilizing a 1 mL syringe pump with a 12-port multiposition valve and an internal magnetic stir bar enables precise mixing, bead dispersion, and magnetic capture for consistent bioconjugate synthesis. The LIS platform achieved a 99.6% bead recovery with 0.4 mg of MIS (1 μm in diameter), outperforming the 83% recovery of manual techniques, and maintained an 83% recovery at reduced scales of 0.2 mg, surpassing manual yields of 76%. As a proof-of-concept, MIS conjugated with anti-SARS-CoV2 antibodies (6 μg/400 mg beads) were synthesized and validated for viral RNA isolation from COVID-19-positive samples, demonstrating high immunocapture efficiency comparable to manual methods but with significantly reduced time and labor requirements. This automated synthesis of antibody-MIS enables the scalable, reproducible production of bioconjugated materials, supporting advanced applications in diagnostic assays, therapeutic delivery, and microfluidic integrations. The LIS approach thus enhances the scope of biomolecular conjugate synthesis, offering streamlined workflows that are suited for downstream analytical and bioanalytical applications. LIS is a versatile, automated system for preparing MIS that researchers can adapt for various targets, particularly when commercial products are unavailable, are prohibitively expensive, or require custom carriers.

摘要

我们展示了使用注射器实验室(LIS)平台首次自动合成磁性免疫吸附剂(MIS)的方法,该方法通过碳二亚胺介导的共价结合促进抗体与磁珠的生物共轭。这种方法是传统手动方法的一种高效、可重复的替代方案,最大限度地减少了移液步骤、涡旋操作和孵育过程,降低了操作偏差。利用配备12端口多位置阀和内部磁力搅拌棒的1 mL注射泵,能够实现精确混合、磁珠分散和磁性捕获,以实现一致的生物共轭物合成。LIS平台使用0.4 mg的MIS(直径1μm)实现了99.6%的磁珠回收率,优于手动技术83%的回收率,并且在0.2 mg的较小规模下保持了83%的回收率,超过了手动方法76%的产率。作为概念验证,合成了与抗SARS-CoV2抗体(6μg/400 mg磁珠)共轭的MIS,并验证了其从COVID-19阳性样本中分离病毒RNA的能力,结果表明其免疫捕获效率与手动方法相当,但时间和劳动力需求显著降低。这种抗体-MIS的自动合成能够实现生物共轭材料的可扩展、可重复生产,支持诊断检测、治疗递送和微流控集成等先进应用。因此,LIS方法扩大了生物分子共轭物合成的范围,提供了适合下游分析和生物分析应用的简化工作流程。LIS是一种用于制备MIS的通用自动化系统,研究人员可以针对各种目标进行调整,特别是在没有商业产品、产品价格过高或需要定制载体的情况下。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eab6/12311662/6039df44aeba/ao4c11126_0001.jpg

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