对接引导相显示用于开发具有新型单链抗体片段（scFv）和碱性磷酸酶的融合蛋白，以进行一步酶联免疫吸附测定（ELISA）检测沙丁胺醇。
Docking guided phase display to develop fusion protein with novel scFv and alkaline phosphatase for one-step ELISA salbutamol detection.
作者信息
Hu Shuai, Yang Guangbo, Chen Zhou, Li Qiuye, Liu Bin, Liu Ming, Zhang Dawei, Chang Shan, Kong Ren
机构信息
Institute of Bioinformatics and Medical Engineering, School of Electrical and Information Engineering, School of Chemical and Environmental Engineering, Jiangsu University of Technology, Changzhou, China.
Beijing New BioConcepts Biotech Co., Ltd., Beijing, China.
出版信息
Front Microbiol. 2023 May 12;14:1190793. doi: 10.3389/fmicb.2023.1190793. eCollection 2023.
INTRODUCTION
Salbutamol (SAL) is a β2 adrenergic receptor agonist which has potential hazardous effects for human health. It is very important to establish a sensitive and convenient method to monitor SAL.
METHODS
Here we introduce a method to combine the information from docking and site specific phage display, with the aim to obtain scFv with high affinity to SAL. First, single chain variable fragment (scFv) antibodies against SAL were generated through phage display. By using molecular docking approach, the complex structure of SAL with antibody was predicted and indicated that H3 and L3 contribute to the binding. Then new libraries were created by randomization specific residues located on H3 and L3 according to the docking results.
RESULTS AND DISCUSSION
Anti-SAL scFv antibodies with high efficiency were finally identified. In addition, the selected scFv was fused with alkaline phosphatase and expressed in to develop a rapid and low-cost one step ELISA to detect SAL.
引言
沙丁胺醇（SAL）是一种β2肾上腺素能受体激动剂，对人类健康有潜在危害。建立一种灵敏便捷的方法来监测SAL非常重要。
方法
在此我们介绍一种将对接信息与位点特异性噬菌体展示相结合的方法，旨在获得对SAL具有高亲和力的单链抗体片段（scFv）。首先，通过噬菌体展示产生抗SAL的单链可变片段（scFv）抗体。利用分子对接方法预测了SAL与抗体的复合物结构，表明H3和L3对结合有贡献。然后根据对接结果，通过对位于H3和L3上的特定残基进行随机化创建新的文库。
结果与讨论
最终鉴定出了高效的抗SAL scFv抗体。此外，将筛选出的scFv与碱性磷酸酶融合并在大肠杆菌中表达，以开发一种快速低成本的一步酶联免疫吸附测定法来检测SAL。
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