IMPROVEMENT OF AN EIA SYSTEM FOR BASIC FIBROBLAST GROWTH-FACTOR BY USE OF BIOTINYLATED ANTIBODY PREPARED WITH NHS-LC-BIOTIN

被引：5
|
作者：
TAKEI, Y [1 ]
HIGASHIRA, H [1 ]
HAYASHI, K [1 ]
机构：
[1] GIFU PHARMACEUT UNIV,DEPT MOLEC BIOL,GIFU 502,JAPAN
来源：
JOURNAL OF CLINICAL LABORATORY ANALYSIS | 1995年 / 9卷 / 02期
关键词：
BREAST CANCER; HIGH MOLECULAR WEIGHT; SENSITIVITY; ISOELECTRIC POINT; SERUM;
D O I：
10.1002/jcla.1860090204
中图分类号：
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号：
1001 ;
摘要：
We improved our previously devised enzyme immunoassay (FIA) system for basic fibroblast growth factor (bFGF) using biotinylated antibody prepared with sulfosuccinimidyl-6-(biotinamido)hexanoate (NHS-LC-Biotin, Pierce), a water-soluble biotin analogue, with an extended spacer arm. The discriminatory detection limit of the improved EIA was found to be 5 pg/ml (0.5 pg/assay tube), sixfold more sensitive than that of the previous system. The reproducibility of within- and between-assay series was 5.10-8.61% and 5.99-8.69%, respectively; and recovery of exogenous bFGF from serum was approximately 102%. Employing the improved EIA system, we investigated by chromatofocusing chromatography the isoelectric points of two immunoreactive bFGFs (high-molecular-weight bFGF-like immunoreactive substance, designated as HMW-bFGF-LI and 16-kd bFGF-LI, having the same molecular weight as recombinant bFGF) detected in serum from a breast cancer patient. As a result, the pi value of HMW-bFGF-LI was estimated to be 7.13, and that of 16 kd bFGF-LI, 9.58. (C) 1995 Wiley-Liss, Inc.
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页码：96 / 100
页数：5
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