M-Protein semi-quantification in MM serum patients based on Immuno-Capture and Liquid Chromatography coupled to High Resolution Mass Spectrometry

Muccio, Stephané; Tavernier, Alexandra; Rouchon, Marie-Claude; Roccon, Alain; Dai, Shujia; Boutet, Valérie; Mouret, Dominique; Pradeilles, Béatrice; Finn, Greg; Macé, Sandrine; DiNoto, Deborah; Fedeli, Olivier

doi:10.1016/j.clml.2019.09.249

Cited by 3 publications

(4 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 31, 32 Consequently, more sensitive and specific assays have been introduced to detect M-protein. 33, 34, 35, 36, 37 An isatuximab-specific antibody-capture assay to mitigate M-protein interference on immunofixation is now Conformité Européenne marked in the European Union, US Food and Drug Administration 510(k) approved, 38 and submitted for regulatory clearance in other regions. 39 However, at the time of the interim analysis of the IKEMA study, the isatuximab-specific assay was not available for use; therefore, M-protein interference was analyzed through high-resolution mass spectrometry, preceded by immuno-capture and liquid chromatography.…”

Section: Discussionmentioning

confidence: 99%

Depth of response and response kinetics of isatuximab plus carfilzomib and dexamethasone in relapsed multiple myeloma

et al. 2022

Self Cite

View full text Add to dashboard Cite

The IKEMA study (Randomized, Open Label, Multicenter Study Assessing the Clinical Benefit of Isatuximab Combined With Carfilzomib [Kyprolis®] and Dexamethasone Versus Carfilzomib With Dexamethasone in Patients With Relapse and/or Refractory Multiple Myeloma Previously Treated With 1 to 3 Prior Lines; #NCT03275285) was a randomized, open-label, multicenter phase 3 study investigating isatuximab plus carfilzomib and dexamethasone (Isa-Kd) vs Kd in patients with relapsed multiple myeloma. This subanalysis analyzed the depth of response of Isa-Kd vs Kd. The primary end point was progression-free survival (PFS); secondary end points included overall response rate, very good partial response or better (≥VGPR) rate, complete response (CR) rate, and minimal residual disease (MRD) negativity rate (assessed in patients with ≥VGPR by next-generation sequencing at a 10−5 sensitivity level). At a median follow-up of 20.7 months, deeper responses were observed in the Isa-Kd arm vs the Kd arm, with ≥VGPR 72.6% vs 56.1% and CR of 39.7% vs 27.6%, respectively. MRD negativity occurred in 53 (29.6%) of 179 patients in the Isa-Kd arm vs 16 (13.0%) of 123 patients in the Kd arm, with 20.1% (Isa-Kd, 36 of 179 patients) vs 10.6% (Kd, 13 of 123 patients) reaching MRD-negative CR status. Achieving MRD negativity resulted in better PFS in both arms. A positive PFS treatment effect was seen with Isa-Kd in both MRD-negative patients (hazard ratio, 0.578; 95% CI, 0.052-6.405) and MRD-positive patients (hazard ratio, 0.670; 95% CI, 0.452-0.993). Exploratory analysis indicates that both current CR and MRD-negative CR rates are underestimated due to M-protein interference (potential adjusted CR rate, 45.8%; potential adjusted MRD-negative CR rate, 24.0%). In conclusion, there was a clinically meaningful improvement in depth of response with Isa-Kd. The CR rate in Isa-Kd was 39.7%. Mass spectrometry suggests that the potential adjusted CR rate could reach an unprecedented 45.8% of patients treated with Isa-Kd.

show abstract

Section: Discussionmentioning

confidence: 99%

Depth of response and response kinetics of isatuximab plus carfilzomib and dexamethasone in relapsed multiple myeloma

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…The IC-LC-HRMS assay allowed differentiation of Isa and M-protein and thus to overcome the interference of Isa with M-protein measurement observed in conventional IFE assays. Most samples that were below the limit of quantification (1000 µg/mL) in SPEP analysis were quantified using LC-HRMS with a 10 µg/mL equivalent alemtuzumab limit of quantification [10].…”

Section: Dear Editormentioning

confidence: 99%

Antibody interference and response kinetics of isatuximab plus pomalidomide and dexamethasone in multiple myeloma

et al. 2021

Self Cite

View full text Add to dashboard Cite

“…3 To eliminate potential isatuximab interference with serum M-protein measurement in patients enrolled in clinical studies, we developed and validated a hybrid assay based on immunocapture and liquid chromatography coupled to highresolution mass spectrometry (IC-HPLC-HRMS). 4 The enhanced selectivity of HRMS allows for a highly sensitive assay method which could be very useful in discriminating between several Igs, based on their specific mass (m/z). In a serum sample from an untreated patient with MM, an abnormal high monoclonal Ig concentration would be characterized by the presence of a "spike" in the polyclonal Ig mass spectra, at a specific mass characteristic of the Mprotein in the patient.…”

Section: ■ Introductionmentioning

confidence: 99%

“…To eliminate potential isatuximab interference with serum M-protein measurement in patients enrolled in clinical studies, we developed and validated a hybrid assay based on immunocapture and liquid chromatography coupled to high-resolution mass spectrometry (IC-HPLC-HRMS) . The enhanced selectivity of HRMS allows for a highly sensitive assay method which could be very useful in discriminating between several Igs, based on their specific mass ( m / z ).…”

Section: Introductionmentioning

confidence: 99%

Validated Method Based on Immunocapture and Liquid Chromatography Coupled to High-Resolution Mass Spectrometry to Eliminate Isatuximab Interference with M-Protein Measurement in Serum

et al. 2021

Self Cite

View full text Add to dashboard Cite

In multiple myeloma (MM) disease, malignant plasma cells produce excessive quantities of a monoclonal immunoglobulin (Ig), known as M-protein. M-protein levels are measured in the serum of patients with MM using electrophoresis techniques to determine the response to treatment. However, therapeutic monoclonal antibodies, such as isatuximab, may confound signals using electrophoresis assays. We developed a robust assay based on immunocapture and liquid chromatography coupled to high-resolution mass spectrometry (IC-HPLC-HRMS) in order to eliminate this interference. Following immunocapture of Ig and free light chains (LC) in serum, heavy chains (HC) and LC were dissociated using dithiothreitol, sorted by liquid chromatography and analyzed using HRMS (Q-Orbitrap). This method allowed the M-proteins to be characterized and the signals from isatuximab and M-proteins to be discriminated. As Mprotein is specific to each patient, no standards were available for absolute quantification. We therefore used alemtuzumab (an IgG kappa mAb) as a surrogate analyte for the semiquantification of M-protein in serum. This assay was successfully validated in terms of selectivity/specificity, accuracy/precision, robustness, dilution linearity, and matrix variability from 10.0 to 200 μg/mL in human serum. This method was used for clinical assessment of samples and eliminated potential interference due to isatuximab when monitoring patients with MM.

show abstract

M-Protein semi-quantification in MM serum patients based on Immuno-Capture and Liquid Chromatography coupled to High Resolution Mass Spectrometry

Cited by 3 publications

References 0 publications

Depth of response and response kinetics of isatuximab plus carfilzomib and dexamethasone in relapsed multiple myeloma

Depth of response and response kinetics of isatuximab plus carfilzomib and dexamethasone in relapsed multiple myeloma

Antibody interference and response kinetics of isatuximab plus pomalidomide and dexamethasone in multiple myeloma

Validated Method Based on Immunocapture and Liquid Chromatography Coupled to High-Resolution Mass Spectrometry to Eliminate Isatuximab Interference with M-Protein Measurement in Serum

Contact Info

Product

Resources

About