Enzyme immunoassay for erythrocyte aldose reductase

Abstract

This two-site immunoassay measures erythrocyte aldose reductase by using monoclonal and polyclonal antibodies to recombinant human enzyme. Total incubation time is 2.5 h, and the limit of detection is < 0.05 microgram/L. Analytical recovery tested with blood samples from healthy and diabetic individuals was 101-106%. Average CVs within and between assays were 3.7% and 4.8%, respectively. The enzyme content determined by this system correlated well with the activity of aldose reductase isolated from the same erythrocyte preparations. The amount of erythrocyte aldose reductase per milligram of hemoglobin was higher in women than in men (P < 0.001), but no significant correlation was observed between the amount of enzyme and the age of the individuals. This assay method should provide useful clinical information to optimize administration of aldose reductase inhibitors for effective prevention and treatment of diabetic complications.

Nanoscale Dielectrophoretic Spectroscopy of Individual Immobilized Mammalian Blood Cells

“…The most abundant protein in the red blood cell membrane is the band three protein, also known as AE1 (23), which transports bicarbonate into and out of the red blood cell. Current methods for the analysis of living blood include antibody testing (24), fluorescence-based optical microscopy, (25) and measurement of blood cell sedimentation time (26). High resolution analysis of blood cells has been performed with freeze fracture atomic force microscopy (27) and electron microscopy (23).…”

Aldose reductase gene polymorphism is associated with progression of diabetic nephropathy in Japanese patients with type 1 diabetes mellitus

“…We examined erythrocytes for AR protein content in eight patients from each group by a two‐site enzyme‐linked immunosorbent assay (ELISA) using monoclonal antibodies directed against recombinant human AR [17]. The mean values were 11.96 ± 2.61 ng/mg Hb in Z+2 homozygotes group, 13.95 ± 2.61 ng/mg Hb in Z+2 heterozygotes group and 15.03 ± 4.87 ng/mg Hb in the no Z+2 group respectively (not significant: p = 0.235).…”

Aldose Reductase as a Key Target in the Prevention and Treatment of Diabetic Retinopathy: A Comprehensive Review

“…An uniquely developed immunoassay technique that uses a particular antibody directed against AR can measure the enzyme's concentration [58]. In a study it was shown to strongly correspond with the activity of AR that was extracted from the same people's erythrocytes [62].…”