Comparative study between liquid phase and solid phase radioimmunoassay system for determination of C-pipted in human serum

Abstract

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.