ENZYME-LINKED ANTIGLOBULIN CONSUMPTION ASSAY (ELACA) : FOR QUANTITATION OF IGG, IGM AND C3 ON THE SURFACE MEMBRANE OF PLATELETS AND RED CELLS

Abstract

In autoimmune disease involving blood cells, quantitation of immunoproteins bound to the surface membrane of the blood cells are of diagnostic importance. Several methodologies are currently available to quantitate the cell bound immunoproteins, however not suited for usage in routine laboratories of developing countries. The purpose of this study is to develop an assay which is sensitive, reproducible, avoids costly reagents and uses reagents which have long shelf life and are readily available in the market. Also an assay that could quantitate IgG, IgM and C3 on the surface of all three types of blood cells. This study chose to develop an ELISA procedure as it has potentials to fulfil the criteria. This study used the method of Gills and Follea to quantitate IgG, IgM and C3 on the surface membrane of platelets as a starting guide, modified and optimised the assay by taking note of several conditions like - dilution of serum coating the well, antibody dilution, cell count and non-specific binding.