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A procedure for producing an anti-AXL nanobody in E. coli

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机构: [1]College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China [2]Laboratory of Quality & Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai, 201306, China [3]State Key Laboratory of Drug Research, The CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China [4]University of Chinese Academy of Sciences, Beijing, 100049, China [5]Drug Discovery Shandong Laboratory, Bohai Rim Advanced Research Institute for Drug Discovery, Yantai, Shandong, 264117, China [6]Shandong Provincial Key Laboratory of Biopharmaceuticals, Shandong Academy of Pharmaceutical Sciences, Jinan, 250101, China [7]Department of Pharmacy, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China [8]National Cancer Center/National Clinical Research Center for Cancer/Hebei Cancer Hospital, Chinese Academy of Medical Sciences, Langfang, 065001, China
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关键词: AXL Nanobody Expression Purification

摘要:
As one of the receptors of the TAM family, AXL plays a vital role in stem cell maintenance, angiogenesis, immune escape of viruses and drug resistance against tumors. In this study, the truncated extracellular segment containing two immunoglobulin-like domains of human AXL (AXL-IG), which has been confirmed to bind growth arrest specific 6 (GAS6) by structural studies [1], was expressed in a prokaryotic expression system and then purified. Immunizing camelid with the purified AXL-IG as antigen could lead to the production of unique nanobodies composed of only variable domain of heavy chain of heavy-chain antibody (VHH), which are around 15 kD and stable. We screened out a nanobody A-LY01 specific binding to AXL-IG. We further determined the affinity of A-LY01 to AXL-IG and revealed that A-LY01 could specifically recognize full-length AXL on the surface of HEK 293T/17 cells. Our study provides appropriate support for the development of diagnostic reagents and antibody therapeutics targeting AXL.Copyright © 2023 Elsevier Inc. All rights reserved.

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出版当年[2023]版:
大类 | 4 区 生物学
小类 | 4 区 生化研究方法 4 区 生化与分子生物学 4 区 生物工程与应用微生物
最新[2025]版:
大类 | 4 区 生物学
小类 | 4 区 生化研究方法 4 区 生化与分子生物学 4 区 生物工程与应用微生物
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出版当年[2023]版:
Q4 BIOCHEMICAL RESEARCH METHODS Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
最新[2023]版:
Q4 BIOCHEMICAL RESEARCH METHODS Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

影响因子: 最新[2023版] 最新五年平均 出版当年[2023版] 出版当年五年平均 出版前一年[2022版]

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第一作者机构: [1]College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China [2]Laboratory of Quality & Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai, 201306, China [3]State Key Laboratory of Drug Research, The CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China
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通讯机构: [1]College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China [2]Laboratory of Quality & Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai, 201306, China [3]State Key Laboratory of Drug Research, The CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China [4]University of Chinese Academy of Sciences, Beijing, 100049, China [7]Department of Pharmacy, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China [8]National Cancer Center/National Clinical Research Center for Cancer/Hebei Cancer Hospital, Chinese Academy of Medical Sciences, Langfang, 065001, China [*1]State Key Laboratory of Drug Research, The CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China. [*2]College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China. [*3]Department of Pharmacy/ National Cancer Center/ National Clinical Research Center for Cancer/ Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China.
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