High-Throughput Therapeutic Antibody Interference-Free High-Resolution Mass Spectrometry Assay for Monitoring M-Proteins in Multiple Myeloma
| Autor: | Naiyu Zheng, Oscar Puig, Craig Titsch, Zheng Ouyang, Renuka Pillutla, Jianing Zeng, Yang J Zhang, Rasa Santockyte |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
medicine.drug_class
Immunoglobulins Antineoplastic Agents 010402 general chemistry Immunoglobulin light chain Mass spectrometry Monoclonal antibody Interference (wave propagation) Antibodies Monoclonal Humanized 01 natural sciences Sensitivity and Specificity Mass Spectrometry Analytical Chemistry medicine Humans Elotuzumab Throughput (business) Multiple myeloma Chromatography biology Chemistry 010401 analytical chemistry medicine.disease 0104 chemical sciences High-Throughput Screening Assays biology.protein Antibody Multiple Myeloma medicine.drug Chromatography Liquid |
| Zdroj: | Analytical chemistry. 93(2) |
| ISSN: | 1520-6882 |
| Popis: | Measurement of monoclonal antibodies (M-proteins) plays an important role in the diagnosis and treatment monitoring of multiple myeloma. Currently available M-protein assays have several limitations, particularly because of their lack of sensitivity and propensity to therapeutic antibody (t-mAb) interference. A previously described mass spectrometry method termed monoclonal immunoglobulin rapid accurate mass measurement (miRAMM) is more sensitive than current clinical tests and can provide a solution for resolving t-mAb interferences. However, the original miRAMM workflow is too complex for the throughput needed to analyze a large number of samples. Here, we describe a high-throughput liquid chromatography-high-resolution mass spectrometry (HT-LC-HRMS) approach that employs a fully automated immunocapture step, significantly improved immunoglobulin recovery, simplified chromatography, and high throughput (HT) data processing. In this HT-LC-HRMS approach, raw spectra of the peaks eluting from the LC column during the predefined time period are automatically deconvoluted without the need to identify and monitor the retention time of each patient-specific M-protein. The deconvoluted peak heights of M-protein and therapeutic antibody light chain are conveniently used for quantitation. With the total LC-HRMS measurement time being only 11.0 min, this method was able to differentiate between the M-protein and elotuzumab mass signatures in 91 out of 92 (98.9%) multiple myeloma serum samples tested. The single interference case was resolved using the mass signature of a heavy chain. In addition to resolving t-mAb interference, the developed assay has a 25-fold improvement in sensitivity over immunofixation electrophoresis and can potentially provide an objective tracking of M-proteins in patients with complete response. |
| Databáze: | OpenAIRE |
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