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使用酶联免疫吸附测定法测定多价受体的解离常数:以展示肌钙蛋白T特异性肽的M13噬菌体为例

Determination of the Dissociation Constant for Polyvalent Receptors Using ELISA: A Case of M13 Phages Displaying Troponin T-Specific Peptides.

作者信息

Machera Sebastian J, Niedziółka-Jönsson Joanna, Jönsson-Niedziółka Martin, Szot-Karpińska Katarzyna

机构信息

Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, Warsaw 01-244, Poland.

出版信息

ACS Omega. 2023 Jul 12;8(29):26253-26262. doi: 10.1021/acsomega.3c02551. eCollection 2023 Jul 25.

Abstract

Phage-derived affinity peptides have become widespread thanks to their easy selection via phage display. Interactions between a target protein and its specific peptide are similar to those between antibodies and antigens. The strength of these non-covalent complexes may be described by the dissociation constant (). In this paper, protein-specific peptides are exposed on the pIII protein present in the M13 bacteriophage virion with up to five copies. Therefore, one phage particle can bind from one to five ligands. Here, we discuss the dependences between phage-displayed peptides and their ligands in solution using a model system based on troponin T (TnT) binding phages. Moreover, a method of calculating values from ELISA experiments was developed and is presented. The determined values are in the picomolar range.

摘要

噬菌体衍生的亲和肽因其通过噬菌体展示易于筛选而广泛应用。靶蛋白与其特异性肽之间的相互作用类似于抗体与抗原之间的相互作用。这些非共价复合物的强度可用解离常数()来描述。在本文中,蛋白质特异性肽暴露于M13噬菌体病毒粒子中存在的pIII蛋白上,拷贝数最多为五个。因此,一个噬菌体颗粒可以结合一到五个配体。在这里,我们使用基于肌钙蛋白T(TnT)结合噬菌体的模型系统,讨论溶液中噬菌体展示肽与其配体之间的相关性。此外,还开发并展示了一种从ELISA实验计算值的方法。所测定的值在皮摩尔范围内。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6880/10373194/5e4c0b9e0a7d/ao3c02551_0002.jpg

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