title: Immunological detection of glutamyl aminopeptidase in urine samples from cisplatin-treated rats.
abstract: OBJECTIVE
  The aim of this work is to demonstrate if urinary excretion of glutamyl aminopeptidase (GluAp) can be quantified by immunological methods.
  
  
  METHODS
  Urine samples from control and cisplatin-treated rats (n = 10 each group) were obtained at 1, 8, and 15 days after cisplatin injection. GluAp was analyzed by kinetic fluorimetry, ELISA, and immunoblotting. Sensitivity and specificity was studied for fluorimetric activity and ELISA 24 h after cisplatin injection. We also analyzed the predictive value over renal dysfunction at the end of the experiment.
  
  
  RESULTS
  GluAp was easily detected by immunoblotting and ELISA, and its urinary excretion was increased in cisplatin-treated rats (p < 0.01). Results obtained with ELISA were strongly correlated (r = 0.8186; p < 0.0001) with fluorimetric activity. Kinetic fluorimetry was the method with the highest AUC (AUC = 1) and the highest predictive value over serum creatinine (r = 0.7630; p = 0.0001) and body weight increase (r = -0.8721; p < 0.0001).
  
  
  CONCLUSIONS
  GluAp can be detected in urine samples with immunological methods, making possible the development of an antibody-based kit for its determination. Its excretion correlates with the extent of renal dysfunction in cisplatin-treated rats, confirming its value as an early marker of renal damage that can be a diagnostic aid in renal diseases.
authors.affiliation: Área de Fisiología, Departamento de Ciencias de la Salud, Universidad de Jaén, Jaén, Spain.
authors.forename: Sebastián
authors.initials: S
authors.lastname: Montoro-Molina
authors.affiliation: Unidad de Nefrología, FIBAO, Hospital Virgen de las Nieves, Granada, Spain.
authors.forename: Andrés
authors.initials: A
authors.lastname: Quesada
authors.affiliation: Área de Fisiología, Departamento de Ciencias de la Salud, Universidad de Jaén, Jaén, Spain.
authors.forename: Piedad V
authors.initials: PV
authors.lastname: Zafra-Ruiz
authors.affiliation: Departamento de Anatomía Patológica, IBIMER, Universidad de Granada, Granada, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: O'Valle
authors.affiliation: Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.
authors.forename: Félix
authors.initials: F
authors.lastname: Vargas
authors.affiliation: Unidad de Nefrología, FIBAO, Hospital Virgen de las Nieves, Granada, Spain.
authors.forename: María Del Carmen
authors.initials: Mdel C
authors.lastname: de Gracia
authors.affiliation: Unidad de Nefrología, FIBAO, Hospital Virgen de las Nieves, Granada, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Osuna
authors.affiliation: Área de Fisiología, Departamento de Ciencias de la Salud, Universidad de Jaén, Jaén, Spain.
authors.forename: Rosemary
authors.initials: R
authors.lastname: Wangensteen
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000633:toxicity*; D015153:Blotting, Western; D002945:Cisplatin / Q000633:toxicity*; D043384:Glutamyl Aminopeptidase / Q000652:urine*; D008297:Male; D012372:ROC Curve; D017208:Rats, Wistar; D051437:Renal Insufficiency / Q000139:chemically induced / Q000201:enzymology / Q000652:urine
chemical_list: D000970:Antineoplastic Agents; D043384:Glutamyl Aminopeptidase; D002945:Cisplatin
keywords: ELISA; Immunoblotting; Nephrotoxicity; Renal dysfunction; Urinary markers