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基于PCR的简化蛋白质定量测定,使用DNA阿普塔默和甲基纤维素作为阻断剂

Oleksij Redcenko¹, Magda Tumova¹, Petr Draber¹

¹Laboratory of Signal Transduction, Institute of Molecular Genetics of the Czech Academy of Sciences, Vídeňská 1083, 142 20 Prague, Czech Republic.

International journal of molecular sciences

|January 11, 2024

概括

这项研究引入了一种简化的一井定量实时聚合酶连锁反应 (qPCR) 试验,使用aptamers. 甲基纤维素有效地阻断了聚烯井中的非特异性结合,简化了基于aptamer的分子量化.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
分析化学分析化学

背景情况:

由于其独特的结构,寡核酸胺为分子结合提供了高亲和力和特异性.
使用aptamers或aptamer-antibody组合的定量实时聚合酶连锁反应 (qPCR) 分析可以量化目标分子.
目前的基于aptamer的qPCR测试通常是复杂的,缺乏可靠性.

研究的目的:

探索在标准聚烯PCR板井内进行完整的基于aptamer的qPCR试验的可行性.
确定有效的阻断剂,以最大限度地减少聚烯井中的非特异性结合.
开发一种简化和可靠的单井qPCR试验,用于分子量化.

主要方法:

研究了蛋白质与聚烯PCR井的结合.
评估了各种阻断剂以防止非特异性结合.
鉴定了甲基纤维素作为一种有效的阻断剂,将其性能与牛血清白蛋白 (BSA) 进行比较.

主要成果:

聚烯井表现出高效的蛋白质结合,使得单井测定性能.
甲基纤维素被证明是一种有效的阻断剂,与BSA相比或优于BSA.
甲基纤维素具有明确组成和非动物来源的优势.

结论:

一个简化的,单井的qPCR试验,利用在聚烯井中固定的aptamers是可行的.
甲基纤维素是BSA的一个合适和有利的替代品,用于阻止这些测试.
这种方法可以在明确的条件下对分子进行敏感和可靠的量化.

关键词:

它们是DNA吸收剂.阻止代理商的阻止代理商.免疫球蛋白是一种免疫球蛋白.甲基纤维素的使用方法聚合酶连锁反应的发生.聚乙烯井井的使用情况

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