采用双抗体酶免疫测定法测定人酰化 ghrelin 血浆浓度时的陷阱。
Pitfalls in the determination of human acylated ghrelin plasma concentrations using a double antibody enzyme immunometric assay.
机构信息
Department of Pediatrics, British Columbia Children's Hospital and Child and Family Research Institute, University of British Columbia, Vancouver, BC, Canada, V6H 3V4.
出版信息
Clin Biochem. 2012 Jan;45(1-2):178-80. doi: 10.1016/j.clinbiochem.2011.10.023. Epub 2011 Nov 7.
OBJECTIVES
To investigate the effect of hemolysis and protease inhibitors on acylated ghrelin (AG) concentrations measured with a double antibody enzyme immunometric assay that uses an acetylcholinesterase (AChE)-Fab' conjugate.
DESIGN AND METHODS
Samples were hemolysed or treated with PHMB (p-hydroxymercuribenzoate), PMSF (phenylmethanesulfonylfluoride) or AEBSF (4-(2-Aminoethyl) benzenesulfonyl fluoride) to prevent AG degradation.
RESULTS
Hemolysis decreased AG concentrations. PHMB or PMSF did not affect the assay. The standard curve was abolished by AEBSF but rescued by addition of a washing step prior to the AChE-Fab' conjugate.
CONCLUSIONS
Hemolysis and AEBSF may affect AG determination.
目的
研究使用乙酰胆碱酯酶（AChE）-Fab' 缀合物的双抗体酶免疫测定法测量酰化生长素释放肽（AG）浓度时，溶血和蛋白酶抑制剂的影响。
设计和方法
对样品进行溶血处理或用 PHMB（对羟基汞苯甲酸）、PMSF（苯甲基磺酰氟）或 AEBSF（4-（2-氨基乙基）苯磺酰氟）处理以防止 AG 降解。
结果
溶血降低了 AG 浓度。PHMB 或 PMSF 不影响测定。AEBSF 使标准曲线失效，但在添加 AChE-Fab' 缀合物之前增加洗涤步骤可使其恢复。
结论
溶血和 AEBSF 可能会影响 AG 的测定。
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