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蛋白质固定方法与纸上的共价键的比较,用于基于纸的酶链免疫吸收剂测定

Yang Chen¹, Kaewta Danchana¹, Takashi Kaneta²

¹Department of Chemistry, Okayama University, 3-1-1 Tsushimanaka, Kita-ku, Okayama, 700-8530, Japan.

Analytical and bioanalytical chemistry

|October 7, 2024

概括

在基于纸张的酶链免疫吸收试验 (P-ELISA) 中,优化纸张上的抗体固定是至关重要的. 与NaIO4相比,APTS-glutaraldehyde方法提高了灵敏度和可重现性,厚纸改善了信号稳定性.

科学领域:

生物技术是生物技术.
分析化学分析化学
材料科学材料科学材料科学

背景情况:

基于纸质的酶链免疫吸收试验 (P-ELISA) 提供了一个低成本的诊断平台.
有效的抗体固定是P-ELISA性能和稳定性的关键.
希夫基形成是表面结合的常见方法.

研究的目的:

为了比较两种方法,在纸上引入基组,以使抗体不动.
在P-ELISA中优化抗体固定和测试性能的条件.
评估纸张厚度和化学减少对试验稳定性和可重复性的影响.

主要方法:

在纸上使用希夫基形成对抗体 (人体IgG) 固定.
通过 sodium periodate (NaIO4) 和 APTS-glutaraldehyde 引入化基.
优化固定,酶反应和还原步骤.
分析测定灵敏度,可重现性和稳定性,使用皮过氧化酶结合抗人类IgG (HRP-anti-IgG) 和TMB基质.

主要成果:

APTS-甲方法表现出比NaIO4方法更高的灵敏度和可重复性.
与NaBH4的胺基键的化学减少对灵敏度和稳定性产生了最小的影响,但降低了可重复性.
发现更厚的纸质基板是首选的,防止洗过程中的扭曲,稳定分析信号.

结论:

在P-ELISA中建议使用APTS-glutaraldehyde方法进行抗体固定,因为它提高了分析性能.
纸张厚度是提高P-ELISA稳定性和可靠性的关键因素.
进一步优化P-ELISA协议可以导致更稳定和敏感的纸质诊断工具.

关键词:

协价结合是一种共价结合.艾丽莎·伊丽莎·伊丽莎固定化固定化固定化基于纸质的酶链接免疫吸收剂测定.蛋白质蛋白质是一种蛋白质.

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