TY - JOUR
T1 - Optimized LC-MS/MS Method for the Detection of ppCCK(21-44)
T2 - A Surrogate to Monitor Human Cholecystokinin Secretion
AU - Foreman, Rachel E.
AU - Miedzybrodzka, Emily L.
AU - Eiríksson, Finnur Freyr
AU - Thorsteinsdóttir, Margrét
AU - Bannon, Christopher
AU - Wheller, Robert
AU - Reimann, Frank
AU - Gribble, Fiona M.
AU - Kay, Richard G.
N1 - Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society.
PY - 2023/9/1
Y1 - 2023/9/1
N2 - The hormone cholecystokinin (CCK) is secreted postprandially from duodenal enteroendocrine cells and circulates in the low picomolar range. Detection of this digestion and appetite-regulating hormone currently relies on the use of immunoassays, many of which suffer from insufficient sensitivity in the physiological range and cross-reactivity problems with gastrin, which circulates at higher plasma concentrations. As an alternative to existing techniques, a liquid chromatography and mass spectrometry-based method was developed to measure CCK-derived peptides in cell culture supernatants. The method was initially applied to organoid studies and was capable of detecting both CCK8 and an N-terminal peptide fragment (prepro) ppCCK(21-44) in supernatants following stimulation. Extraction optimization was performed using statistical modeling software, enabling a quantitative LC-MS/MS method for ppCCK(21-44) capable of detecting this peptide in the low pM range in human plasma and secretion buffer solutions. Plasma samples from healthy individuals receiving a standardized meal (Ensure) after an overnight fast were analyzed; however, the method only had sensitivity to detect ppCCK(21-44). Secretion studies employing human intestinal organoids and meal studies in healthy volunteers confirmed that ppCCK(21-44) is a suitable surrogate analyte for measuring the release of CCK in vitro and in vivo.
AB - The hormone cholecystokinin (CCK) is secreted postprandially from duodenal enteroendocrine cells and circulates in the low picomolar range. Detection of this digestion and appetite-regulating hormone currently relies on the use of immunoassays, many of which suffer from insufficient sensitivity in the physiological range and cross-reactivity problems with gastrin, which circulates at higher plasma concentrations. As an alternative to existing techniques, a liquid chromatography and mass spectrometry-based method was developed to measure CCK-derived peptides in cell culture supernatants. The method was initially applied to organoid studies and was capable of detecting both CCK8 and an N-terminal peptide fragment (prepro) ppCCK(21-44) in supernatants following stimulation. Extraction optimization was performed using statistical modeling software, enabling a quantitative LC-MS/MS method for ppCCK(21-44) capable of detecting this peptide in the low pM range in human plasma and secretion buffer solutions. Plasma samples from healthy individuals receiving a standardized meal (Ensure) after an overnight fast were analyzed; however, the method only had sensitivity to detect ppCCK(21-44). Secretion studies employing human intestinal organoids and meal studies in healthy volunteers confirmed that ppCCK(21-44) is a suitable surrogate analyte for measuring the release of CCK in vitro and in vivo.
KW - Biological Transport
KW - Bodily Secretions
KW - Cholecystokinin
KW - Chromatography, Liquid
KW - Humans
KW - Tandem Mass Spectrometry
UR - http://www.scopus.com/inward/record.url?scp=85169032476&partnerID=8YFLogxK
U2 - 10.1021/acs.jproteome.3c00272
DO - 10.1021/acs.jproteome.3c00272
M3 - Article
C2 - 37591880
AN - SCOPUS:85169032476
SN - 1535-3893
VL - 22
SP - 2950
EP - 2958
JO - Journal of Proteome Research
JF - Journal of Proteome Research
IS - 9
ER -