DEVELOPMENT OF AN AUTOMATED FLOW-INJECTION CHEMILUMINESCENCE IMMUNOASSAY FOR HUMAN-IMMUNOGLOBULIN-G

The development of a competitive solid phase flow-injection immunoassay with on-column chemiluminescence detection is described. The immunoreactor column consists of a transparent teflon tubing packed with immobilized antibodies. It is placed in front of the photomultiplier tube. The entire assay takes place in this immunoreactor cell. The assay is performed by injection of a mixture of sample HIgG and acridinium labeled HIgG followed by hydrogen peroxide to initiate the chemiluminescence reaction. One assay cycle including regeneration takes 7 min. The system is fully automated and controlled by computer to ensure exact timing resulting in a precision of 1 to 3%, dependent on concentration. The detection limit was 7 fmol. The method was applied to serum samples without any sample clean-up. The average recovery from spiked serum samples was 103%.