液相和固相放射免疫分析系统测定人血清中C-pipted的比较研究

Ahmed Abdelsadek, Prof. Khaled Sallam, Mahmoued eltawoosy
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引用次数: 0

摘要

本研究旨在检验用于定量人类血清中C肽的固相和液相放射免疫分析(RIA)系统的优缺点。基本试剂已经生产出来。第一个RIA是125I-C肽示踪剂。一种间接示踪剂制备利用氯胺-T和酪氨酸甲酯(TME)。使用HPLC柱,从反应混合物中分离出示踪剂。第二种试剂是先前在另一项工作中产生的多克隆抗体。使用高度纯化的多克隆抗体进行包被管。使用高度纯化的C肽创建了一组C肽标准品。建立了液相系统放射免疫分析法(RIA)测定人血清中C肽水平。为了进行这项研究,研究了许多参数,包括交叉反应、特异性和非特异性结合、样品体积、温度影响、孵育时间和有效期。使用固相和液相检测不同的人血清样品。统计研究显示,这两种手术的结果之间有很强的相关性,但差异很小。固相比液相具有许多优点。观察到高非特异性结合和低特异性结合百分比。相反,高结合率和低成本是液相系统的主要优点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparative study between liquid phase and solid phase radioimmunoassay system for determination of C-pipted in human serum
This study aimed to examine the benefits and drawbacks of both solid phase and liquid phase radioimmunoassay (RIA) systems used to quantify C-peptide in human serum. The fundamental reagents were produced. The first RIA was 125 I-C-peptide tracer. An indirect tracer preparation utilized chloramine-T and tyrosine methyl ester (TME). Using HPLC column, the tracer was separated from the reaction mixture. The second reagent was the polyclonal antibody which was produced previously in another work. The coated tube was carried out using a highly purified polyclonal antibody. A set of C-peptide standards was created using highly purified C-peptide. A liquid phase system radioimmunoassay (RIA) was created to measure the C-peptide levels in human serum. Numerous parameters were investigated to perform this study including cross reaction, specific and nonspecific binding, sample volume, temperature influence, incubation time, and the valid period. Different samples of human serum were examined using both solid and liquid phases. The statistical study showed a strong relation between the outcomes of the two procedures with minor differences. The solid phase has many advantages over the liquid phase. A high nonspecific binding and a low specific binding percent were observed. On the contrary a high binding percent and the low cost are the main advantages in Liquid phase system.
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