3D Printed Polylactic Acid (PLA) Well Plates for Enzyme Inhibition Studies: The Case of Pancreatic Lipase

H. Stamatis, D. Gournis, A. Polydera, Stamatia Spyrou, Konstantinos Spyrou, Elena Gkantzou, Anastasia Skonta
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引用次数: 2

Abstract

3D printed PLA has already been demonstrated for several biotechnological applications, including enzymes immobilization. The prerequisites for an efficient screening assay include using small volumes of reagents, low cost, and rapid screening of large numbers of compounds and extracts. Hence, assays based on microtiter plates are predominant. Thus, designing and fabricating scaffolds on a similar scale, which could serve as immobilization carriers, and their recruitment in inhibitors screening studies is of great significance, adding both enzyme stability and reuse potentiality of the biocatalytic system in assay merits. In this work, pancreatic lipase was immobilized on 3D-printed PLA microwells for enzyme inhibitor screening. XPS analysis demonstrated the successful modification of the PLA scaffolds. The immobilized enzyme displayed high levels of operational, thermal, and storage stability under the tested conditions. The IC50 values for PPL inhibition were calculated for Orlistat, a model lipase inhibitor, and olive leaf extract, a promising natural compound. This is the first study reporting the use of 3D-printed PLA wells with an immobilized enzyme for inhibitor screening assay.
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3D打印聚乳酸孔板用于酶抑制研究:以胰脂肪酶为例
3D打印PLA已经被证明用于几种生物技术应用,包括酶固定。高效筛选试验的先决条件包括使用小体积试剂、低成本和快速筛选大量化合物和提取物。因此,以微量滴度板为基础的检测方法占主导地位。因此,设计和制造类似规模的可作为固定化载体的支架及其在抑制剂筛选研究中的招募具有重要意义,增加了生物催化体系在酶稳定性和重复使用潜力方面的优点。本研究将胰脂肪酶固定在3d打印的PLA微孔上进行酶抑制剂筛选。XPS分析证实了PLA支架的成功修饰。在测试条件下,固定化酶显示出高水平的操作、热和储存稳定性。计算了脂肪酶抑制剂奥利司他和天然化合物橄榄叶提取物对PPL抑制作用的IC50值。这是第一个使用3d打印PLA孔和固定化酶进行抑制剂筛选试验的研究。
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