Establishment of Chemiluminescent Immunoassay for Determination of Carcinoembrynic Antigen

Objective To establish a method of chemiluminescent immunoassay for determination of carcinoembrynic antigen(CEA) in human serum. Methods The two-site enzyme immunoassay was based on the direct sandwich technique.A monoclonal antibody was bonded to the microplate for coating and the conjugation of alkaline phosphatase to another polyclonal antibody was performed by NaIO4 method.Chemiluminescent immunoassay was established by optimizing CSPD and SapphireII luminescent system.The reference range was defined by testing 800 serum samples from healthy subjects.The comparison for 100 serum samples from patients was assayed by the bayer Acs:180 Automated Immunoassay System. Results The detectable minimum was 0.5 ng/mL.The linear scope was from 0.5 to 640.0 ng/mL.Average inter and intra-assays were 8.6% and 7.5%,respectively.The recoveries ranged from 97.4% to 101.8%.The cross-reacting rate for AFP(3 μg/mL),Ferritin(10 μg/mL),HAS(200 g/L),IgG(100 g/L) were≤0.015%,0.034%,4.5×10-8 and 5.0×10-8,respectively.Compared with bayer Acs:180 Automated Immunoassay System,the relative coefficient was 0.968(P0.001). Conclusion A successful establishment of chemiluminescent immunoassay provides a way for clinical determination of CEA.