Multianalyte immunoassay chip for detection of tumor markers by chemiluminescent and colorimetric methods

Most cancers developed an elevation of the level of at least two markers associated with their incidence. Simultaneous detection of multi-tumor markers associated with a particular type of cancer plays an important role in cancer diagnostic. Here, a multianalyte immunoassay chip for simple and sensitive detection of tumor markers with chemiluminescent and colorimetric methods was proposed, in which carcinoembryonic antigen (CEA) and carbohydrate antigen (CA19-9) that associated with colorectal cancer were detected as model. The immunoassay chip was fabricated by co-immobilization of CEA/CA19-9 antibody on a glass slide with gamma-glycidoxypropyltrimethoxysilane as linkage. Through sandwiched immunoreactions, CEA, CA19-9, and their corresponding enzyme tracers, alkaline phosphatase-labeled anti-CEA and horseradish peroxidase-labeled anti-CA19-9, were introduced on the chip. Then, they were sequentially detected by chemiluminescent method in the range of 0.5-80 mu g/L and 0.5-80 kU/L with the detection limits of 0.41 mu g/L and 0.36 kU/L at 3 sigma for CEA and CA19-9, respectively. They could also be detected by colorimetric method in the range of 1-200 mu g/L and 5-200 kU/L with the detection limits of 0.25 mu g/L and 1.25 kU/L at 3 sigma for CEA and CA19-9, respectively. All these results demonstrated that the present work provided a promising analytical method for tumor markers' analysis with the advantages of simple analytical procedure, small sample volume and lower cost, which made the proposed method potential for high-throughput detection.