表面等离子共振
检出限
乙型肝炎表面抗原
毕赤酵母
重组DNA
抗原
化学
分析物
色谱法
生物传感器
分子生物学
线性范围
抗体
免疫分析
表面等离子体子
材料科学
乙型肝炎病毒
病毒学
纳米颗粒
生物
生物化学
等离子体子
纳米技术
免疫学
光电子学
病毒
基因
作者
Yew Joon Tam,Zeenathul Nazariah Allaudin,Morvarid Akhavan Rezaei,Nazahah Mustafa,Mohd Azmi Mohd Lila,Abdul Rani Bahaman,Sewn Cen Lo,Joo Shun Tan,Homayoun Hani,Rasedee Abdullah
摘要
Abstract Limit of detection (LOD), limit of quantification, and the dynamic range of detection of hepatitis B surface antigen antibody (anti‐HBs) using a surface plasmon resonance (SPR) chip‐based approach with Pichia pastoris ‐derived recombinant hepatitis B surface antigen (HBsAg) as recognition element were established through the scouting for optimal conditions for the improvement of immobilization efficiency and in the use of optimal regeneration buffer. Recombinant HBsAg was immobilized onto the sensor surface of a CM5 chip at a concentration of 150 mg/L in sodium acetate buffer at pH 4 with added 0.6% Triton X‐100. A regeneration solution of 20 mM HCl was optimally found to effectively unbind analytes from the ligand, thus allowing for multiple screening cycles. A dynamic range of detection of ∼0.00098–0.25 mg/L was obtained, and a sevenfold higher LOD, as well as a twofold increase in coefficient of variance of the replicated results, was shown as compared with enzyme‐linked immunosorbent assay (ELISA). Evaluation of the assay for specificity showed no cross‐reactivity with other antibodies tested. The ability of SPR chip‐based assay and ELISA to detect anti‐HBs in human serum was comparable, indicating that the SPR chip‐based assay with its multiple screening capacity has greater advantage over ELISA.
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