2. Academic Validation
  3. Quantification of anti-SARS-CoV-2 antibodies in human serum with LC-QTOF-MS

Quantification of anti-SARS-CoV-2 antibodies in human serum with LC-QTOF-MS

  • J Pharm Biomed Anal. 2021 Oct 25;205:114319. doi: 10.1016/j.jpba.2021.114319.
Karen A M de Jong 1,  Hilde Rosing 2,  Marit Vermunt 2,  Alwin D R Huitema 3,  Jos H Beijnen 4
Affiliations

Affiliations

  • 1 Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands. Electronic address: Ka.d.jong@nki.nl.
  • 2 Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • 3 Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Clinical Pharmacy, University Medical Center Utrecht, Utrecht, The Netherlands; Department of Pharmacology, Princess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands.
  • 4 Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands; Division of Pharmacoepidemiology and Clinical Pharmacology, Faculty of Science, Department of Pharmaceutical Sciences, Utrecht University, The Netherlands.
PMID: 34416552 DOI: 10.1016/j.jpba.2021.114319
Abstract

The aim of this study was to develop the first quantitative serological test for anti-SARS-CoV-2 Antibodies in human serum with liquid chromatography - quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Other assays, mostly immunoassays, are only qualitative or semi-quantitative, and hence, actual antibody concentrations after SARS-CoV-2 Infection are unknown. In our assay, anti-SARS-CoV-2 Antibodies were isolated with spike protein subunit 1 (S1) coupled to Magnetic Beads. IgG1 signature peptide GPSVFPLAPSSK was selected for quantification using ipilimumab calibration standards and SILuMAb K1 as the stable-isotope labeled internal standard. The anti-SARS-CoV-2 IgG1 calibration range was from 1.35 to 135 nM. Inter-assay accuracies were between 98.8%- 107% with inter-assay precisions between 8.37%- 13.5% measured at 3 concentration levels on three separate occasions. Anti-SARS-CoV-2 IgG1 Antibodies were quantified in PCR-positive patients with mild to severe symptoms. IgM signature peptide DGFFGVPR was detected in patients that recently recovered from COVID-19. A unique and quantitative LC-QTOF-MS method to quantify anti-SARS-CoV-2 IgG1 in serum was successfully developed and its clinical applicability has been demonstrated.

Keywords

Anti-SARS-CoV-2 antibodies; COVID-19; LC-QTOF-MS; Mass spectrometry; SARS-CoV-2; Spike protein.

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