Aptamer based peptide enrichment for quantitative analysis of gonadotropin-releasing hormone by LC-MS/MS

Talanta
S L RichardsM J Raftery

Abstract

Over recent years threats to racing have expanded to include naturally occurring biological molecules, such as peptides and proteins, and their synthetic analogues. Traditionally, antibodies have been used to enable detection of these compounds as they allow purification and concentration of the analyte of interest. The rapid expansion of peptide-based therapeutics necessitates a similarly rapid development of suitable antibodies or other means of enrichment. Potential alternative enrichment strategies include the use of aptamers, which offer the significant advantage of chemical synthesis once the nucleic acid sequence is known. A method was developed for the enrichment, detection and quantitation of gonadotropin-releasing hormone (GnRH) in equine urine using aptamer-based enrichment and LC-MS/MS. The method achieved comparable limits of detection (1 pg/mL) and quantification (2.5 pg/mL) to previously published antibody-based enrichment methods. The intra- and inter-assay precision achieved was less than 10% at both 5 and 20 pg/mL, and displayed a working dynamic range of 2.5-100 pg/mL. Significant matrix enhancement (170 ± 8%) and low analytical recovery (29 ± 15%) was observed, although the use of an isotopically heavy label...Continue Reading

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Citations

Sep 4, 2016·Talanta·Yubin ZhouTerry W J Steele
Jan 15, 2017·Drug Testing and Analysis·A G FragkakiM Tsivou
Mar 16, 2018·Expert Review of Proteomics·Brianna KimAlessandra Luchini
Mar 3, 2017·Clinical Chemistry·Geoffrey S Baird, Andrew N Hoofnagle
May 29, 2020·Biomédica : revista del Instituto Nacional de Salud·Juan David Ospina
Oct 11, 2020·International Journal of Molecular Sciences·Chioma Izzi-EngbeayaWaljit S Dhillo
Jan 14, 2021·Drug Testing and Analysis·Fernando MoreiraMaria de Lourdes Bastos
Sep 2, 2021·Journal of Neuroscience Methods·Taylor D PobranMari L DeMarco

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