Polyethylene Glycol-Mediated Directional Conjugation of Biological Molecules for Enhanced Immunoassays at the Point-of-Care.
Polyethylene Glycol-Mediated Directional Conjugation of Biological Molecules for Enhanced Immunoassays at the Point-of-Care.
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DOI：
10.3390/polym15153316
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发表时间：
2023-08-05
期刊：
影响因子：
5
通讯作者：
Draz MS
中科院分区：
文献类型：
--
作者：
Battalapalli D;Chakraborty P;Jain D;Obaro SK;Gurkan UA;Bonomo RA;Draz MS
快速可靠的即时护理 (POC) 诊断测试可以对全球健康产生重大影响。开发 POC 系统最常见的方法之一是使用特定目标的生物分子。然而，生物分子的缀合可能导致活性降低，这可能会损害所开发系统的分析性能和准确性。为了克服这一挑战，我们提出了一种基于聚合物的交联方案，用于生物分子的受控和定向缀合。我们的方案利用双功能硫醇-聚乙二醇 (PEG)-酰肼聚合物来实现 IgG 抗体与丝网印刷金属电极表面的定点缀合。电极的金属表面首先用硫醇化 PEG 分子进行修饰，使酰肼基团可与氧化 IgG 抗体 Fc 片段中的醛基反应。使用抗肺炎克雷伯菌碳青霉烯酶-2 (KPC-2) 抗体作为用于抗菌素耐药性 (AMR) 测试的模型抗体，我们的结果表明，与标准的基于 N-羟基琥珀酰亚胺 (NHS) 的缀合化学相比，抗体偶联增加了约 10 倍，并且在修饰电极表面有效捕获 (>94%) 目标 KPC-2 酶抗原。这种直接且易于执行的定点抗体偶联策略可用于偶联 POC 测试和开发中常用的其他基于蛋白质和非蛋白质的生物分子，从而增强提高诊断准确性和性能的潜力。
Rapid and reliable point-of-care (POC) diagnostic tests can have a significant impact on global health. One of the most common approaches for developing POC systems is the use of target-specific biomolecules. However, the conjugation of biomolecules can result in decreased activity, which may compromise the analytical performance and accuracy of the developed systems. To overcome this challenge, we present a polymer-based cross-linking protocol for controlled and directed conjugation of biological molecules. Our protocol utilizes a bifunctional thiol-polyethylene glycol (PEG)-hydrazide polymer to enable site-directed conjugation of IgG antibodies to the surface of screen-printed metal electrodes. The metal surface of the electrodes is first modified with thiolated PEG molecules, leaving the hydrazide groups available to react with the aldehyde group in the Fc fragments of the oxidized IgG antibodies. Using anti-Klebsiella pneumoniae carbapenemase-2 (KPC-2) antibody as a model antibody used for antimicrobial resistance (AMR) testing, our results demonstrate a ~10-fold increase in antibody coupling compared with the standard N-hydroxysuccinimide (NHS)-based conjugation chemistry and effective capture (>94%) of the target KPC-2 enzyme antigen on the surface of modified electrodes. This straightforward and easy-to-perform strategy of site-directed antibody conjugation can be engineered for coupling other protein- and non-protein-based biological molecules commonly used in POC testing and development, thus enhancing the potential for improved diagnostic accuracy and performance.
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影响因子：
17.1
作者：
Draz MS;Lakshminaraasimulu NK;Krishnakumar S;Battalapalli D;Vasan A;Kanakasabapathy MK;Sreeram A;Kallakuri S;Thirumalaraju P;Li Y;Hua S;Yu XG;Kuritzkes DR;Shafiee H
通讯作者：
Shafiee H
影响因子：
4.1
作者：
Huang, Chao;Wen, Tian;Jiao, Yong-Jun
通讯作者：
Jiao, Yong-Jun
影响因子：
17.1
作者：
Draz, Mohamed Shehata;Fang, Binbin Amanda;Chen, Fanqing Frank
通讯作者：
Chen, Fanqing Frank
影响因子：
7.4
作者：
Ho, Ja-an Annie;Chang, Heng-Chia;Chou, Chien
通讯作者：
Chou, Chien
影响因子：
19
作者：
Draz, Mohamed Shehata;Moazeni, Maryam;Shafiee, Hadi
通讯作者：
Shafiee, Hadi