An enzyme-based assay for the detection of carboxyethyl lysine-protein adducts - PROJECT SUMMARY
Advanced glycation endproducts (AGEs) are hallmarks of mammalian aging and disease.
Nε-carboxyethyl-lysine (CEL) in particular, is among the most abundant AGEs accumulating in tissue proteins
with aging, and is an important marker of glycation, oxidative stress, and disease. Current techniques for
reliable measurement of CEL are slow, expensive (>$120/sample), and not readily available to many academic
research labs nor easily applied for routine analysis of clinical samples. Development of a convenient method
for measurement of CEL would democratize the research tools used to study AGEs and advance the tools
available to study and treat diseases at the interface of aging, glycation and oxidative stres.
Revel has discovered a CEL oxidase enzyme which oxidizes CEL to lysine. The newly discovered CEL
oxidase provides an opportunity to develop enzymes for diagnostic use in monitoring CEL-modifications. The
goal of this Phase I SBIR is to develop a rapid, one-pot, inexpensive enzymatic test (<$10/sample) to measure
CEL. The proposed assay will be developed analogous to the enzymatic HbA1c test which is used to assess
patient blood sugar levels and diagnose diabetes. In Phase II, we will develop a workflow for CEL detection in
patient samples. Eventually, further engineering of the enzyme for removal of CEL from endogenous proteins
could yield a ground-breaking tool for reversing glycoxidatively damaged proteins in clinical medicine.
