M-inSight®

Ultra sensitive, Personalized MRD testing on Blood

M-inSight® is a personalized, serum-based, Targeted Mass Spectrometry assay for Minimal Residual Disease (MRD) monitoring in Multiple Myeloma. M-inSight® tracks very low level of M-protein,the biomarker secreted by the cancer cells.
M-inSight® Highlights

M-inSight® can see what others can’t

Personalized
blood-based assay

M-inSight® tracks patient-specific clonotypic peptides from the M-protein secreted by the tumor cells.

Ultra-sensitivity is reached by analyzing unique peptides derived from the variable region of the M-protein (clonotypic peptides) corresponding to patient specific V(D)J-gene rearrangements and somatically hypermutated gene sequences.

Benefits compared to MRD - bone marrow testing

M-inSight® on blood is at least as sensitive as the latest generation of MRD evaluation performed on bone marrow and has similar prognostic value in patients with MM.
Bone marrow aspirate is an invasive procedure, not suitable for dynamic MRD monitoring due to limited timepoints. In addition, there is a risk of false negative results with BM-MRD evaluation due to tumor heterogeneity while M-protein quantitation on blood reflects the disease burden.

See deeper than other
M-protein measurements

Routine M-protein diagnostic tests (SPEP, IFE) are reliable and fast but not sensitive enough to detect low M-protein concentrations once the patient is treated.
Intact protein quantitation by Mass Spectrometry (MALDI) has been developed for diagnostic use only as the sensitivity is limited by the polyclonal background.
Because many patients who obtain sCR will eventually relapse, more sensitive assays capable of measuring minimal residual disease (MRD) are urgently needed.

The Workflow

How does M-inSight® work?

M-inSight® works in 2 steps :
  • Patient Definition Assay : M-protein is sequenced from serum, then the best clonotypic peptides are selected from this sequence. This is a one-time process.
  • Patient Follow-up Assessment : M-protein quantitation is obtained over time measuring clonotypic peptides in serum.

 

 
Once the serum sample is sent to our laboratory, we will process the sample and prepare a personalized report that will be released in 3-5 days (1-2 weeks for the Patient Definition Assay).
M-inSight® RESULTS

Low level disease tracking over time

M-inSight® will produce a personalized report, providing physicians important information about the kinetics of the disease and the depth of response to treatment.
 

 

M-inSight® can monitor sustained MRD negativity.

M-inSight® can detect progression far earlier than other current methods.*

The Technology

Ultra sensitive Mass Spectrometry assay to quantify M-Protein

M-inSight® uses a combination of 3 dimensions to reach the highest sensitivity :

High resolution separation by using nano Ultra High Performance Liquid Chromatography (nUPLC) + High resolution mass analyzer with the Orbitrap technology to accurately quantify at the lowest level + Unique Clonotypic peptides targeting avoids the interference with the polyclonal background and therapeutic monoclonal antibody.

 

The main advantages are : higher sensitivity with accurate quantitation, faster throughput, deeper coverage.

 

MS TECHNOLOGY

Different Mass spectrometry assays, different levels of sensitivity

Because  each immunoglobulin has a specific amino-acid sequence and therefore a specific molecular mass, M-protein can be detected by Mass Spectrometry. M-protein’s sequence is unique to each MM patient and it remains unchanged through the different stages of the disease, which makes it a surrogate marker for the presence of clonal plasma cells. M-protein can be detected by MS as a whole (Intact protein) with low sensitivity or as unique fragments (clonotypic peptides) with high sensitivity.

Mass spectrometry

Mass spectrometry is an analytical tool measuring the mass-to-charge ratio (m/z) of one or more molecules present in a sample. These measurements can often be used to calculate the exact molecular weight of the sample components as well. Typically, mass spectrometers can be used to identify unknown compounds via molecular weight determination, to quantify known compounds, and to determine structure and chemical properties of molecules.
How does a mass spectrometer perform such a feat? Every mass spectrometer consists of at least these three components:
Ionization Source , Mass Analyzer, Ion Detection System
Depending on the use and the type of analytes, each component might differ. Intact M-protein is often quantified by MALDI-TOF (Low resolution) while peptide is quantified by ESI-Q-Orbi (High resolution).

Intact Protein MS, MALDI-TOF

In this technique, the intact protein is analyzed on the mass spectrometer (MALDI). Inside the instrument, a laser makes it fly away from the sample to the detector. The time it takes for the protein to reach the detector will depend on its mass. However, IgG’s with similar masses might mask the presence of the M-protein. MALDI MS is often used for high thoughput. However, the low resolution of the mass spectrometer with a lack of structural information decrease the sensitivity of the method. The interference with the polyclonal background has a high negative impact on sensitivity when detecting Intact M-protein by Maldi Mass spectrometry.

Targeted MS, Clonotypic peptides MS

A targeted approach allows a much more sensitive detection and quantitation of the M-protein. The protein is initially digested into small peptides from which the primary structure of the protein is inferred via De novo sequencing. On a further step, clonotypic peptides that have a unique sequence are chosen to quantify the M-protein. Targeted MS fragmentation technology allows to distinguish the M-protein peptide from any other molecule with a similar mass since the MS fragmentation pattern generated for each peptide is sequence dependent, minimizing any possible interference.

To further improve the sensitivity, M-inSight uses an Orbitrap Mass Spectrometer that combines two mass analyzers: a quadrupole mass analyzer for high selectivity and high ion transmission and an Orbitrap mass analyzer for high dynamic range and high-resolution measurements. By combining these mass analyzers and orchestrating the coordination of ions transfer within the instrument, high performance is achieved to unlock new insights into biology and disease mechanisms.