High prevalence of immuno-unreactive intact albumin in urine of diabetic patients.
BACKGROUND
Intact albumin in urine may exist in two forms, immunoreactive and immuno-unreactive. Previous estimates of albuminuria in diabetic urine have only detected immunoreactive forms.


METHODS
High performance liquid chromatography (HPLC) was used in this study to measure both forms of intact albumin (termed total intact albumin) to provide a more accurate measurement of albuminuria compared with radioimmunoassay (RIA) on 97 fresh urine samples from patients with diabetes. Eighty-six control urine samples from volunteers without diabetes were also tested.


RESULTS
There was no significant difference between the two methods for nondiabetic controls. For diabetic urine samples, 91.6% of samples showed a greater concentration of albumin measured by HPLC than RIA. For normoalbuminuric diabetic samples, HPLC gave a mean albumin excretion rate of 12.5 +/- 4.4 microgram/min (SD), whereas RIA gave a rate of 8.0 +/- 6.7 microgram/min (P = 0.004; N = 28). For microalbuminuric samples, there also was a statistically significant difference: HPLC albumin excretion rate, 82.0 +/- 49.9 microgram/min, and RIA, 49.0 +/- 34.6 microgram/min (P = 0.004; N = 30). Thirty-two urine samples were normoalbuminuric by RIA (albumin, 11.4 +/- 3.9 microgram/min), but in the microalbuminuric range as determined by HPLC (albumin, 38.5 +/- 14.4 microgram/min). For urine samples in the macroalbuminuric range, there was no statistically significant difference between HPLC and RIA. Immuno-unreactive albumin was confirmed as albumin, analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption ionization mass spectrometry.


CONCLUSIONS
These studies show that to determine microalbuminuria accurately, there is a need to assess urinary total intact albumin, rather than simply immunoreactive albumin. Am J Kidney Dis 41:336-342.
Comper
Wayne D
WD
Department of Biochemistry and Molecular Biology, Monash University, Clayton; and the Endocrine Unit, Austin & Repatriation Medical Center, Heidelberg, Victoria, Australia.
Osicka
Tanya M
TM
Jerums
George
G
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000418:Albumins / Q000032:analysis / Q000276:immunology*; D000419:Albuminuria / Q000453:epidemiology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003922:Diabetes Mellitus, Type 1 / Q000652:urine*; D003924:Diabetes Mellitus, Type 2 / Q000652:urine*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D015995:Prevalence; D011863:Radioimmunoassay / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D000418:Albumins