高等学校化学学报2026，Vol.47Issue(6)：72-80,9.DOI:10.7503/cjcu20260057
血清蛋白冠对金纳米探针抗体固定策略及其抗原捕获能力的影响
Influence of Serum Protein Corona on Antibody Immobilization Strategies and Antigen Capture Capabilities of Gold Nanoparticle Probes
摘要
Abstract
When nanoprobes are applied for the detection in complex biological matrices such as serum,a protein corona is prone to form on their surface,which may mask the recognition molecules and thus lead to probe inactivation.To investigate the effects of different antibody immobilization strategies on antibody bioactivity and anti-interference performance,gold nanoparticles were used as the carrier in this study to systematically compare three immobilization methods for rabbit anti-mouse IgG:direct physical adsorption,covalent conjugation via mercaptopropionic acid,and site-directed immobilization mediated by staphylococcal protein A.We comprehensively adopted nanoparticle tracking analysis,BCA protein quantitation and dot blot assay to quantitatively evaluate the antibody loading capacity per nanoparticle and the accessible binding orientation of antibodies for each method.Furthermore,a serum-mimicking environment was constructed to analyze the masking effect of protein corona on antibody exposure and the actual target antigen capture capability of the nanoprobes.The experimental results demonstrated that the physical adsorption method via direct co-incubation exhibited the optimal performance in antibody loading capacity and binding orientation optimization.Moreover,this method maintained the highest antibody accessibility and antigen-binding capacity after incubation in serum,and its overall performance was significantly superior to that of the two chemical modification methods.This study reveals that the simple and direct physical adsorption method possesses unique advantages in the preparation of nanoprobes for detection in complex biological samples,which provides novel experimental evidence and practical strategies for optimizing nanoprobe design and improving the reliability of in vitro diagnostics.关键词
金纳米探针/蛋白冠/抗体固定策略/抗体取向/血清检测Key words
Gold nanoprobe/Protein corona/Antibody immobilization strategy/Antibody orientation/Serum detection分类
化学化工引用本文复制引用
石明凡,张释心,仁青央宗,吴琼,陈芳芳..血清蛋白冠对金纳米探针抗体固定策略及其抗原捕获能力的影响[J].高等学校化学学报,2026,47(6):72-80,9.基金项目
国家重点研发计划项目(批准号:2024YFA0918600)、国家自然科学基金(批准号:32271446)和吉林省科技发展计划项目(批准号:YDZJ202601ZYTS395)和2025年吉林大学中日联谊医院MED+X交叉学科培育项目(批准号:2025023)资助.Supported by the National Key Research and Development Program of China(No.2024YFA0918600),the National Nature Science Foundation of China(No.32271446),the Science and Technology Development Project of Jilin Province,China(No.YDZJ202601ZYTS395)and MED+X Inter-disciplinary Incubation Program of Jilin University China-Japan Union Hospital,China(No.2025023). （批准号:2024YFA0918600）
