The objective of this Phase II project is to develop simple, specific and reliable immunoassays to measure the amount of non-enzymatically glycated hemoglobin (glycohemoglobin in human blood). The circulating level of glycohemoglobin is an index of the ambient glucose concentration over the preceding 2-3 months. The assays use an ELISA construct and employ monoclonal antibodies that specifically recognize glycated hemoglobin. Phase I established the feasibility of creating highly specific monoclonal antibodies to glycated hemoglobin and of applying these antibodies in solid phase colorimetric ELISAs for quantitation of glycohemoglobin in human blood. Further modifications will give tests that are easy to use, and give rapid quantitative results. Two types of formats will be developed and validated. In the first type, modification of conventional ELISA system will provide laboratory tests that can be quantitated with an ELISA reader. The second type of format, envisioned for use in the physician office, will allow visual discrimination of color intensity to determine defined levels of glycohemoglobin, or can be used with a novel electronic reader device to give absolute values. The electronic reader device is being developed independently and a working prototype has been assembled. An integrated approach will be used to simplify the manner in which the assays are run, to make them practical for laboratory use and robust for physician office use, to further validate the modified assays, to select the final solid phase matrices and the assay constructs from the combined optima of stability and shelf life of the reagents and the safety and simplicity for technician and/or physician office use.
