McCormick A N, Leach M E, Savidge G, Alhaq A
The Haemophilia Centre, St Thomas' Hospital, London, UK.
Clin Lab Haematol. 2004 Feb;26(1):57-64. doi: 10.1111/j.0141-9854.2003.00572.x.
Surface plasmon resonance was employed to establish a quantitative assay for recombinant FVIII (rFVIII) products using rFVIII as standard. The anti-FVIII monoclonal antibody ESH4 was immobilized onto a carboxymethyldextran surface. A range of rFVIII concentrations were injected over the surface and the binding response enhanced by the addition of a further monoclonal antibody ESH8. Validation using National Institute of Biological Standards and Controls (NIBSC) sixth International rFVIII concentrate standard gave inter- and intra-assay coefficient of variations (CVs) of 7.5 and 3.68% respectively for ESH4-rFVIII binding alone. Enhancement of the binding signal by secondary addition of ESH8 produced inter- and intra-assay CVs of 2.75 and 1.5%. The ESH4 immobilized chip was found to retain binding capacity following regeneration for at least 75 cycles. The assay was found to be unsuitable for quantitation of plasma derived FVIII product but may prove useful for monitoring of rFVIII production.
采用表面等离子体共振技术，以重组凝血因子VIII（rFVIII）为标准品，建立了一种针对rFVIII产品的定量分析方法。抗FVIII单克隆抗体ESH4固定在羧甲基葡聚糖表面。将一系列rFVIII浓度的样品注入该表面，并通过添加另一种单克隆抗体ESH8来增强结合反应。使用英国国家生物标准与控制研究所（NIBSC）第六代国际rFVIII浓缩标准品进行验证，仅针对ESH4-rFVIII结合时，分析间和分析内变异系数（CV）分别为7.5%和3.68%。通过二次添加ESH8增强结合信号后，分析间和分析内CV分别为2.75%和1.5%。发现固定有ESH4的芯片在再生至少75个循环后仍保留结合能力。该分析方法不适用于定量血浆源性FVIII产品，但可能对监测rFVIII的生产有用。
