内蒙古农业大学学报（自然科学版）2026，Vol.47Issue(4)：63-71,9.DOI:10.16853/j.cnki.1009-3575.2026.04.008
HER2特异性的驼源纳米抗体制备与筛选
Preparation and Screening of HER2-Specific Camel-derived Nanobodies
摘要
Abstract
To prepare HER2-specific nanobodies and provide novel candidate molecules for the targeted therapy of HER2-positive tu-mors.Bioinformatics methods were employed to predict the hydrophobicity,signal peptides,transmembrane structure,and second-ary/tertiary structures of the extracellular domain II of HER2.RNA was extracted from 239T cells,and the gene of this region was cloned by RT-PCR.A prokaryotic expression vector was constructed,and the purified protein was used to immunize camels.RNA was extracted from the blood of immunized camels,and high-throughput sequencing was performed directly on the VHH genes of an-tibodies.Nanobodies were detected using Elisa and SDS-PAGE,and gel blocks of appropriate size were subjected to the protein mass spectrometry sequencing.Based on mass spectrometry results,the top five high-abundance nanobodies were selected and sub-jected for molecular docking analysis.The recombinant protein of the extracellular domain II of HER2 was successfully cloned and expressed,with the serum titer of immunized camels reaching as high as 1∶640 000.After high-throughput sequencing and combin-ing the mass spectrometry results,five high-abundance nanobodies(OUT-2334,OUT-8933,OUT-4166,OUT-2966,OUT-2853)were screened.Molecular docking analysis indicated that these nanobodies could all bind to the key amino acid residues in the extra-cellular domain II of HER2,providing new candidate molecules for the targeted therapy of HER2-positive tumors and offering an effi-cient method for nanobody screening.关键词
HER2/纳米抗体/VHH基因/高通量测序/分子对接Key words
HER2/Nanobody/VHH gene/High-throughput sequencing/Molecular docking分类
生物科学引用本文复制引用
黄贞魁,李村院,李晓悦,韩林,倪伟,胡圣伟..HER2特异性的驼源纳米抗体制备与筛选[J].内蒙古农业大学学报（自然科学版）,2026,47(4):63-71,9.基金项目
天池英才项目(CZ001614,CZ001618) （CZ001614,CZ001618）
兵团自然科学支持计划项目(2024DA031) （2024DA031）
石河子大学科研计划项目(JCYJ202313,CXPY202317) （JCYJ202313,CXPY202317）
