Establishment of biotin-streptavidin time-resolved fluoroimmunoassay method for the measurement of heparanase
10.3760/cma.j.issn.2095-2848.2014.04.010
- VernacularTitle:乙酰肝素酶生物素-链亲和素系统时间分辨荧光免疫分析法的建立及应用
- Author:
Bao ZHU
;
Guoqiang XIE
;
Hualong XIAO
;
Biao HUANG
;
Kejing SHAO
;
Yafeng XU
;
Yi ZHANG
- Publication Type:Journal Article
- Keywords:
Fluoroimmunoassay;
Biotin;
Streptavidin;
Heparanase
- From:
Chinese Journal of Nuclear Medicine and Molecular Imaging
2014;34(4):308-311
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a novel TRFIA for the measurement of heparanase (HPA) in serum samples,and investigate its clinical application.Methods The micro-pore plate wells were first coated with partially recombinant murine anti-human HPA monoclonal antibody.Biotin-labeled recombinant HPA protein was then used to compete with HPA in serum samples,and the prepared europium (III)-labeled streptavidin (Eu3+-SA) was used as signal readout for establishing the BSA-TRFIA assay.Using this assay,the serum HPA levels in healthy subjects (n=32) and tumor patients (n=54) were measured.The results of BSA-TRFIA were compared with those of ELISA.Two-sample t test (or t' test),and linear correlation analysis were used to analyze the data.Results The sensitivity of BSA-TRFIA for measuring HPA was 0.33 ug/L.The CV values for intra-batch and inter-batch were 5.29% and 7.54%,respectively.The average recovery rate was 105.5%.The standard curve range was 0-1 000 ug/L.The serum HPA level measured by the BSA-TRFIA method in healthy subjects was (2.03_+ 1.47) Iug/L.In tumor patients,the HPA level was significantly higher:(22.13_+7.38) ug/L (t'=19.388,P
