Internally referenced ImmunoConcentration assays

We have developed a method for incorporating an internal reference zone in addition to the test zone in a single immunoConcentration device. We have illustrated the method by constructing an internally referenced assay for human choriogonadotropin (HCG) in serum. The internally referenced assay, which can be performed in about 5 min, provides a measured overall precision (CV) of approximately 15% at a concentration, in serum, of approximately 50 int. units/L. The test zone consists of an anti-HCG antibody adsorbed onto latex microspheres and entrapped within a porous support. The reference zone contains antibody against alkaline phosphatase (EC 3.1.3.1), which has also been adsorbed onto latex microspheres and trapped within the support. The use of an internal reference moderates the effects of variations in assay conditions on the interpretation of assay results.