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Development of a sandwich ELISA to detect circulating, soluble IRAP as a potential disease biomarker.
Vear, Anika; Thalmann, Claudia; Youngs, Kristina; Hannan, Natalie; Gaspari, Tracey; Chai, Siew Yeen.
Afiliación
  • Vear A; Department of Physiology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3800, Australia.
  • Thalmann C; Department of Physiology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3800, Australia.
  • Youngs K; Department of Physiology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3800, Australia.
  • Hannan N; Department of Obstetrics and Gynaecology, University of Melbourne, Heidelberg, VIC, 3084, Australia.
  • Gaspari T; Mercy Perinatal, Mercy Hospital for Women, Heidelberg, VIC, 3084, Australia.
  • Chai SY; Department of Pharmacology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3800, Australia.
Sci Rep ; 13(1): 17565, 2023 11 24.
Article en En | MEDLINE | ID: mdl-38001104
There is growing interest in the use of the enzyme, insulin regulated aminopeptidase (IRAP), as a biomarker for conditions such as cardio-metabolic diseases and ischemic stroke, with upregulation in its tissue expression in these conditions. However, quantification of circulating IRAP has been hampered by difficulties in detecting release of the truncated, soluble form of this enzyme into the blood stream. The current study aimed to develop a sandwich ELISA using novel antibodies directed towards the soluble portion of IRAP (sIRAP), to improve accuracy in detection and quantification of low levels of sIRAP in plasma. A series of novel anti-IRAP antibodies were developed and found to be highly specific for sIRAP in Western blots. A sandwich ELISA was then optimised using two distinct antibody combinations to detect sIRAP in the low nanogram range (16-500 ng/ml) with a sensitivity of 9 ng/ml and intra-assay variability < 10%. Importantly, the clinical validity of the ELISA was verified by the detection of significant increases in the levels of sIRAP throughout gestation in plasma samples from pregnant women. The specific and sensitive sandwich ELISA described in this study has the potential to advance the development of IRAP as a biomarker for certain diseases.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Aminopeptidasas / Insulina Límite: Female / Humans / Pregnancy Idioma: En Revista: Sci Rep Año: 2023 Tipo del documento: Article País de afiliación: Australia Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Aminopeptidasas / Insulina Límite: Female / Humans / Pregnancy Idioma: En Revista: Sci Rep Año: 2023 Tipo del documento: Article País de afiliación: Australia Pais de publicación: Reino Unido