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Plasma AVP is a valuable biomarker for the diagnosis of central diabetes insipidus (CDI) and is commonly measured using radioimmunoassay (RIA). However, RIA has several drawbacks, including a long hands-on time, complex procedures, and handling of radioisotopes with special equipment and facilities. In this study, we developed a bioassay to measure plasma AVP levels using HEK293 cells expressing an\u00a0engineered V2R and a cAMP biosensor. To achieve high sensitivity, we screened V2R orthologs from 11 various mammalian species and found that the\u00a0platypus V2R (pV2R) responded to AVP with approximately six-fold higher sensitivity than that observed by the\u00a0human V2R. Furthermore, to reduce cross-reactivity with desmopressin (DDAVP), a V2R agonist used for CDI treatment, we introduced a previously described point mutation into pV2R, yielding an approximately 20-fold reduction of\u00a0responsiveness to DDAVP\u00a0while maintaining responsiveness\u00a0to\u00a0AVP. Finally, a comparison of plasma samples from 12 healthy individuals demonstrated a strong\u00a0correlation (Pearson's correlation value: 0.90) between our bioassay and RIA. Overall, our assay offers a more rapid and convenient method for quantifying plasma AVP concentrations than existing techniques.<\/jats:p>","DOI":"10.1038\/s41598-024-60035-4","type":"journal-article","created":{"date-parts":[[2024,4,24]],"date-time":"2024-04-24T15:02:01Z","timestamp":1713970921000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A cAMP-biosensor-based assay for measuring plasma arginine\u2013vasopressin levels"],"prefix":"10.1038","volume":"14","author":[{"given":"Kosuke","family":"Doi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kouki","family":"Kawakami","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatsuya","family":"Ikuta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asuka","family":"Inoue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,4,24]]},"reference":[{"key":"60035_CR1","doi-asserted-by":"publisher","first-page":"599","DOI":"10.1097\/00000542-200609000-00026","volume":"105","author":"TA Treschan","year":"2006","unstructured":"Treschan, T. 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