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1953 Implementation of a Mass Spectroscopic Method (Mass-Fix) to Replace Traditional Electrophoresis Methods for Patients with Monoclonal Gammopathies

Program: Oral and Poster Abstracts
Session: 653. Multiple Myeloma: Clinical and Epidemiological: Poster I
Hematology Disease Topics & Pathways:
Assays, Emerging technologies, Technology and Procedures, Pathology, Serologic Tests
Saturday, December 7, 2024, 5:30 PM-7:30 PM

David Murray, MD, PhD1*, Mindy Kohlhagen, BS1*, Jon Coker, PhD2*, Maria Alice V. Willrich, PhD, MSc3*, Surendra Dasari, PhD4* and Angela Dispenzieri, MD5

1Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN
2Xylo Technologies, Mayo Clinic, Rochester, MN
3Department of Laboratory Medicine, Mayo Clinic, Rochester, MN
4Department of Quantitative Health Sciences, Mayo Clinic, Rochester, MN
5Division of Hematology, Mayo Clinic, Rochester, MN

Background

The detection, isotyping and quantitation of over-expressed immunoglobulin(s) (Igs) (M-protein) is key for diagnosing and monitoring patients with monoclonal gammopathies. Traditionally, our lab has used a combination of agarose gel protein electrophoresis (SPEP) and immunofixation electrophoresis (IFE) for these purposes. In 2018, a mass spectrometric method (Mass-Fix) was implemented to replace IFE for M-protein isotyping. This work documents the validation of a quantitative version (Q-Mass-Fix) to replace both SPEP and IFE.

Methods

24,645 samples submitted for M-protein testing between 7/2018 and 9/2023 with quantitative IgG, IgA and IgM measurements, Mass-Fix analysis and SPEP results were utilized for this study. A separate cohort of 100,000 M-protein negative samples were utilized to mathematically model the spectral shapes IgG, IgA and IgM polyclonal background. To detect M-proteins, software was developed that fits the polyclonal Ig portion of the spectra and M-proteins are detected as peaks above the polyclonal background. M-proteins isotypes are determined by matching similar peaks in spectra in 5 sperate immuno enrichments (IgG, IgA, IgM, kappa and lambda). M-protein quantitation was obtained by multiplying total M-protein isotype quantity by the fraction area of the M-protein in the spectra. M-protein dilutional studies were used to examine the linearity and level of detection of Q-Mass-Fix.

Results

SPEP and Q-Mass-Fix M-protein quantitation had the best agreement between the range of 10 to 40 g/L. Below 10 g/L, Q-Mass Fix had on average, lower M-protein values. Above 40 g/dL, Q-Mass-Fix values were on average, higher than SPEP. The automated isotyping assignment was 98.8 percent consistent with the isotype assigned by visual inspection. Dilutional studies revealed that Q-Mass-Fix quantitation was linear over a range of 70 to 0.04 g/L (slope 1.0, r2 0.99) with an analytical limit of quantitation of 0.1 g/L and a limit of detection of 0.04 g/L.

Conclusion

Q-Mass-Fix represents a significant improvement in comparison to our current SPEP method. Q-Mass-Fix is more capable of removing the interference of polyclonal Igs below 10 g/L and is not subjective to the dye saturation effect at high M-protein levels. The increased analytical measuring range and improved limit of detection of detection could be beneficial in tracking patient’s response. Q-Mass Fix is more automated than SPEP and the workflow in our laboratory.

Disclosures: Murray: The Binding Site: Patents & Royalties: Intellectual Property Rignts Licensed with potential royalties. Dasari: The Binding Site: Patents & Royalties: Intellectual Property Rights licensed to Binding Site with potential royalties. Dispenzieri: BMS: Consultancy, Research Funding; Alnylam: Research Funding; Takeda: Consultancy, Research Funding; Pfizer: Research Funding; Janssen: Research Funding; Alexion: Consultancy, Research Funding; HaemaloiX: Research Funding.

*signifies non-member of ASH