Antibody-Mediated Screening of Peptide Inhibitors for Monoamine Oxidase-B (MAO-B) from an Autodisplayed FV Library

化学 模拟电影 单克隆抗体 分子生物学 塞莱吉林 抗体 单胺氧化酶B 生物化学 噬菌体展示 单胺氧化酶 生物 免疫学 病理 医学 疾病 帕金森病
作者
Jeong Soo Sung,Ji-Hong Bong,Tae Gyeong Yun,Yeonju Han,Yusun Park,Jaeyong Jung,Soo Jeong Lee,Min‐Jung Kang,Joachim Jose,Misu Lee,Jae‐Chul Pyun
出处
期刊:Bioconjugate Chemistry [American Chemical Society]
卷期号:33 (6): 1166-1178 被引量:11
标识
DOI:10.1021/acs.bioconjchem.2c00107
摘要

Inhibitors for monoamine oxidase-B (MAO-B) were screened from an FV library with a randomized complementarity-determining region 3 (CDR3) region using a monoclonal antibody against dopamine. As the first step, the FV library was expressed on the outer membrane of E. coli by site-directed mutagenesis of the randomized CDR3 region. Among the FV library, variants with a binding affinity to monoclonal antibodies against dopamine were screened and cloned. From the comparison of the binding activity of the screened clones to a control clone with a modified FV antibody (only with CDR1 and CDR2), the CDR3 regions of screened clones were determined to directly interact with the monoclonal antibody against dopamine. These CDR3 sequences were then synthesized as mimotopes (mimicking peptides) of dopamine. The inhibitory activity of two mimotopes against MAO-B was analyzed using HeLa cells overexpressing MAO-B, as well as using activated human astrocytes; their inhibitory activity was compared to that of a commercial inhibitor of MAO-B, selegiline. The inhibition efficiency of the two mimotopes (in comparison with selegiline) was estimated to be 67.2% and 69.4% in the HeLa cells and 64.4% and 58.0% in the human astrocytes. The gene expression pattern in astrocytes after treatment with the two mimotopes was also analyzed and compared with that in the human astrocytes treated with selegiline. Finally, the interaction between two mimotopes and MAO-B was analyzed using docking simulation, and the candidate regions of MAO-B for the interaction with each mimotope were explored through the docking simulation.

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