Measurement of urinary annexin V by ELISA and its significance as a new urinary-marker of kidney disease

• Published in: Clinica chimica acta 2000-08, Vol.298 (1), p.29-43
• Main Authors: Matsuda, Ryuko, Kaneko, Noboru, Horikawa, Yoshifumi, Chiwaki, Fumiko, Shinozaki, Makoto, Abe, Shichiro, Yumura, Wako, Nihei, Hiroshi, Ieiri, Tamio
• Format: Article
• Language: English
• Subjects: Adult; Aged; Annexin A5 - urine; Biological and medical sciences; Biomarkers - urine; Blotting, Western; Diabetes Mellitus - urine; Drug Stability; ELISA; Enzyme-Linked Immunosorbent Assay; Female; Glomerulonephritis, IGA - urine; Humans; Hydrogen-Ion Concentration; Hypertension - urine; Investigative techniques, diagnostic techniques (general aspects); Kidney Diseases - urine; Kidney Failure, Chronic - urine; Lupus Erythematosus, Systemic - urine; Male; Medical sciences; Middle Aged; Monoclonal antibody; Myocardial Ischemia - urine; Nephrotic syndrome; Nephrotic Syndrome - urine; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Proteinuria - urine; Reference Values; Urinary annexin V; Urinary system
• ISSN: 0009-8981; 1873-3492
• DOI: 10.1016/S0009-8981(00)00218-7

To confirm the significance of excretion of annexin V into the urine and the change of urinary annexin V concentration in kidney disease, a sandwich enzyme-linked immunosorbent assay (ELISA) was developed using two monoclonal antibodies. Urinary annexin V concentration was measured in healthy individuals and patients with kidney and other diseases. Urinary annexin V did not change over a range of pH between 5.0 and 8.0, and was stable during the course of the study for 24 h at room temperature and for 8 days at 4°C. The mean urinary annexin V concentration in 105 normal healthy individuals was 1.5±1.5 ng/ml, while that in patients with nephrotic syndrome and systemic lupus erythematosis (SLE) nephritis was 9.3±9.1 and 6.6±6.7 ng/ml, respectively, and that in IgA nephropathy and chronic renal failure was 2.6±2.1 and 1.3±0.7 ng/ml, respectively. Annexin level correlated with urinary protein concentration ( r=0.717), but not the serum creatinine concentration, blood urea nitrogen (BUN) and 24-h creatinine clearance. Mean urinary annexin V concentration in patients with ischemic heart disease, hypertension, and diabetes mellitus was 1.4±1.0, 1.4±1.1, and 1.7±1.3 ng/ml, respectively. In one case of relapsing nephrotic syndrome, the urinary annexin V concentration was markedly increased in the early phase after admission and then decreased. This patient later required hemodialysis. These results suggest that a high urinary annexin V concentration may be an indicator of acute renal injury related to the urinary protein level.