Abstract

A new protein detector of 2,4,6-trinitrophenol (TNP, picric acid) is concisely reported herein for the first time. In brief, the gene of a specific scFv fragment, namely 3.5, was fused separately with the gene of β-lactamase and, subsequently, expressed in the periplasmic space of Escherichia coli, affording in such a way the recombinant fusion protein 3.5-scFv-β-lactamase. This bifunctional immunoreagent tool was further convincingly demonstrated for being capable to directly detect trinitrophenol-tris(hydroxymethyl)aminomethane (TNP-Tris), a less toxic TNP derivative of choice compared to TNP itself, with competitive sensitivity (50 ± 2 fmol or 175 ± 6 pg/mL).

摘要

本文首次简要报道了一种新型的2,4,6-三硝基苯酚（TNP，苦味酸）蛋白质检测器。简而言之，将特定scFv片段的基因，即3.5，与β-内酰胺酶基因分开融合，随后在大肠杆菌的周质空间中表达，以这种方式提供重组融合蛋白3.5-scFv- β-内酰胺酶。这种双功能免疫试剂工具进一步令人信服地证明能够直接检测三硝基苯酚-三(羟甲基)氨基甲烷 (TNP-Tris)，与 TNP 本身相比，这是一种毒性较小的 TNP 衍生物，具有竞争灵敏度（50 ± 2 fmol 或 175 ± 6 皮克/毫升）。