Gábor Transform-Based Antibody Quantitation in Serum: An Interlaboratory Liquid Chromatography/High-Resolution Mass Spectrometry Investigation DOI

Kayd L. Meldrum,

Andrew K. Swansiger,

Jacob Koscho

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(43), P. 17413 - 17422

Published: Oct. 16, 2024

Therapeutic monoclonal antibodies (t-mAbs) are crucial for treating various conditions, including cancers and autoimmune disorders. Accurate quantitation pharmacokinetic monitoring of t-mAbs in serum essential, but current methods like ligand binding assays (LBAs) bottom-up peptide liquid chromatography-tandem mass spectrometry (LC-MS/MS) can lack the sensitivity specificity needed to meet clinical demands. Emerging techniques using high-resolution (HRMS) top-down middle-up approaches offer improved ability accurately quantify mAb proteoforms apart from degradation products by keeping sample proteins intact or minimizing digestion. This study describes first use Gábor transform (GT)-based iFAMS Quant+ software a t-mAb (vedolizumab) ∼400 samples an Agilent 6545XT AdvanceBio Q-TOF at University Oregon. These results compared previously validated laboratory-developed test (LDT) Mayo Clinic utilizing Thermo Q Exactive Plus Orbitrap. The method used conventional extracted ion chromatograms (XICs) select charge states quantitation, while utilized GT-based state deconvolution, background subtraction, signal integration. Calibration quality control (QC) analyses Passing-Bablok regression 351 subject demonstrated excellent agreement between two methods. workflow exhibited unique advantages characterizing interferents analyte anomalies due its deconvolution-based approach.

Language: Английский

Gábor Transform-Based Antibody Quantitation in Serum: An Interlaboratory Liquid Chromatography/High-Resolution Mass Spectrometry Investigation DOI

Kayd L. Meldrum,

Andrew K. Swansiger,

Jacob Koscho

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(43), P. 17413 - 17422

Published: Oct. 16, 2024

Therapeutic monoclonal antibodies (t-mAbs) are crucial for treating various conditions, including cancers and autoimmune disorders. Accurate quantitation pharmacokinetic monitoring of t-mAbs in serum essential, but current methods like ligand binding assays (LBAs) bottom-up peptide liquid chromatography-tandem mass spectrometry (LC-MS/MS) can lack the sensitivity specificity needed to meet clinical demands. Emerging techniques using high-resolution (HRMS) top-down middle-up approaches offer improved ability accurately quantify mAb proteoforms apart from degradation products by keeping sample proteins intact or minimizing digestion. This study describes first use Gábor transform (GT)-based iFAMS Quant+ software a t-mAb (vedolizumab) ∼400 samples an Agilent 6545XT AdvanceBio Q-TOF at University Oregon. These results compared previously validated laboratory-developed test (LDT) Mayo Clinic utilizing Thermo Q Exactive Plus Orbitrap. The method used conventional extracted ion chromatograms (XICs) select charge states quantitation, while utilized GT-based state deconvolution, background subtraction, signal integration. Calibration quality control (QC) analyses Passing-Bablok regression 351 subject demonstrated excellent agreement between two methods. workflow exhibited unique advantages characterizing interferents analyte anomalies due its deconvolution-based approach.

Language: Английский

Citations

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