Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus
Current immunoassay techniques for analyzing clinically relevant parathyroid hormone (PTH) circulating fragments cannot distinguish microheterogeneity among structurally similar molecular species. This hinders the identification of molecular species and the capture of target analyte information. Sin...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-10-01
|
| Series: | Proteomes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2227-7382/12/4/30 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850239256866324480 |
|---|---|
| author | Hicham Benabdelkamel Refat M. Nimer Afshan Masood Maha Al Mogren Anas M. Abdel Rahman Assim A. Alfadda |
| author_facet | Hicham Benabdelkamel Refat M. Nimer Afshan Masood Maha Al Mogren Anas M. Abdel Rahman Assim A. Alfadda |
| author_sort | Hicham Benabdelkamel |
| collection | DOAJ |
| description | Current immunoassay techniques for analyzing clinically relevant parathyroid hormone (PTH) circulating fragments cannot distinguish microheterogeneity among structurally similar molecular species. This hinders the identification of molecular species and the capture of target analyte information. Since structural modifications are important in disease pathways, mass spectrometry can detect, identify, and quantify heterogeneous ligands captured by antibodies. We aimed to create a sensitive and selective multiple reaction monitoring–mass spectrometric immunoassay analysis (MRM-MSIA)-based method for detecting and quantifying PTH fragments or proteoforms for clinical research. Our study established MRM transitions using triple-quadrupole tandem mass spectrometry for the signature peptides of five PTH fragments. This method was validated according to FDA guidelines, employing the mass spectrometric immunoassay (MSIA) protocol to bolster detection selectivity and sensitivity. This validated approach was applied by analyzing samples from type 2 diabetes mellitus (T2DM) patients with and without vitamin D deficiency. We found serum PTH fragments associated with vitamin D deficiency in patients with and without T2DM. We developed and validated the MRM-MSIA technique specifically designed for the detection and quantification (amino acid (aa38–44), (aa45–51), and (aa65–75)) of these fragments associated with vitamin D deficiency and T2DM. This study is the first to accurately quantify plasma PTH fragments using MRM-MSIA, demonstrating its potential for clinical diagnostics. |
| format | Article |
| id | doaj-art-399b489d69e74bbdb5e562ebde5bd8fc |
| institution | OA Journals |
| issn | 2227-7382 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Proteomes |
| spelling | doaj-art-399b489d69e74bbdb5e562ebde5bd8fc2025-08-20T02:01:13ZengMDPI AGProteomes2227-73822024-10-011243010.3390/proteomes12040030Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes MellitusHicham Benabdelkamel0Refat M. Nimer1Afshan Masood2Maha Al Mogren3Anas M. Abdel Rahman4Assim A. Alfadda5Proteomics Resource Unit, Obesity Research Center, College of Medicine, King Saud University, Riyadh 11451, Saudi ArabiaDepartment of Medical Laboratory Sciences, Jordan University of Science and Technology, Irbid 22110, JordanProteomics Resource Unit, Obesity Research Center, College of Medicine, King Saud University, Riyadh 11451, Saudi ArabiaMetabolomics Section, Department of Clinical Genomics, Center for Genome Medicine, King Faisal Specialist Hospital and Research Centre (KFSHRC), Riyadh 11211, Saudi ArabiaMetabolomics Section, Department of Clinical Genomics, Center for Genome Medicine, King Faisal Specialist Hospital and Research Centre (KFSHRC), Riyadh 11211, Saudi ArabiaProteomics Resource Unit, Obesity Research Center, College of Medicine, King Saud University, Riyadh 11451, Saudi ArabiaCurrent immunoassay techniques for analyzing clinically relevant parathyroid hormone (PTH) circulating fragments cannot distinguish microheterogeneity among structurally similar molecular species. This hinders the identification of molecular species and the capture of target analyte information. Since structural modifications are important in disease pathways, mass spectrometry can detect, identify, and quantify heterogeneous ligands captured by antibodies. We aimed to create a sensitive and selective multiple reaction monitoring–mass spectrometric immunoassay analysis (MRM-MSIA)-based method for detecting and quantifying PTH fragments or proteoforms for clinical research. Our study established MRM transitions using triple-quadrupole tandem mass spectrometry for the signature peptides of five PTH fragments. This method was validated according to FDA guidelines, employing the mass spectrometric immunoassay (MSIA) protocol to bolster detection selectivity and sensitivity. This validated approach was applied by analyzing samples from type 2 diabetes mellitus (T2DM) patients with and without vitamin D deficiency. We found serum PTH fragments associated with vitamin D deficiency in patients with and without T2DM. We developed and validated the MRM-MSIA technique specifically designed for the detection and quantification (amino acid (aa38–44), (aa45–51), and (aa65–75)) of these fragments associated with vitamin D deficiency and T2DM. This study is the first to accurately quantify plasma PTH fragments using MRM-MSIA, demonstrating its potential for clinical diagnostics.https://www.mdpi.com/2227-7382/12/4/30diabetesvitamin D deficiencymass spectrometric immunoassay (MSIA)multiple reaction monitoringparathyroid hormone |
| spellingShingle | Hicham Benabdelkamel Refat M. Nimer Afshan Masood Maha Al Mogren Anas M. Abdel Rahman Assim A. Alfadda Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus Proteomes diabetes vitamin D deficiency mass spectrometric immunoassay (MSIA) multiple reaction monitoring parathyroid hormone |
| title | Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus |
| title_full | Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus |
| title_fullStr | Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus |
| title_full_unstemmed | Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus |
| title_short | Multiple Reaction Monitoring–Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus |
| title_sort | multiple reaction monitoring mass spectrometric immunoassay analysis of parathyroid hormone fragments with vitamin d deficiency in patients with diabetes mellitus |
| topic | diabetes vitamin D deficiency mass spectrometric immunoassay (MSIA) multiple reaction monitoring parathyroid hormone |
| url | https://www.mdpi.com/2227-7382/12/4/30 |
| work_keys_str_mv | AT hichambenabdelkamel multiplereactionmonitoringmassspectrometricimmunoassayanalysisofparathyroidhormonefragmentswithvitaminddeficiencyinpatientswithdiabetesmellitus AT refatmnimer multiplereactionmonitoringmassspectrometricimmunoassayanalysisofparathyroidhormonefragmentswithvitaminddeficiencyinpatientswithdiabetesmellitus AT afshanmasood multiplereactionmonitoringmassspectrometricimmunoassayanalysisofparathyroidhormonefragmentswithvitaminddeficiencyinpatientswithdiabetesmellitus AT mahaalmogren multiplereactionmonitoringmassspectrometricimmunoassayanalysisofparathyroidhormonefragmentswithvitaminddeficiencyinpatientswithdiabetesmellitus AT anasmabdelrahman multiplereactionmonitoringmassspectrometricimmunoassayanalysisofparathyroidhormonefragmentswithvitaminddeficiencyinpatientswithdiabetesmellitus AT assimaalfadda multiplereactionmonitoringmassspectrometricimmunoassayanalysisofparathyroidhormonefragmentswithvitaminddeficiencyinpatientswithdiabetesmellitus |