MASSterclass™

Low abundance protein biomarker assay development and screening

There is a growing recognition of the gap that lies between biomarker discovery and the final clinical assay. Discovery platforms are designed to analyse potentially thousands of analytes in parallel in a limited but well defined set of samples, resulting in a hit-list of candidate biomarkers. To prove clinical utility and performance these candidate biomarkers need to be screened in larger sample cohorts and therefore require rapid development of multiplex protein assays. Conventional validation assays, such as ELISAs, chip and solution-based multiplexed assays and western blots, require antibodies that are often unavailable or of poor quality. The process of generating antibodies is time consuming and costly and is therefore not a practical solution for screening the hit-list of proteins derived from discovery.

Coupled with MASStermind^®, our highly-sensitive proteomics biomarker discovery engine we have developed MASSterclass™, a targeted mass spectrometry based antibody-free platform for multiplexed protein quantitation. The platform employs a novel pre-fractionation scheme coupled with a targeted triple quadrupole MS read-out (based on multiple reaction monitoring or MRM) capable of reaching limits of quantitation down to the low pg/mL level.

Key strengths of the MASSterclass™ platform for protein quantitation:
Protein assay development of up to 50 proteins in 8 weeks No need for antibodies Accurate protein quantitation up to 10 pg/ml in plasma Only 25 µl of sample volume required for quantitation of all proteins in a single assay Analytes unambiguously identified by the mass spectrometer Detects peptide surrogates of the protein targets (rather than the intact proteins) so less subject to variability due to sample degradation Build assays to regions of protein sequence that are homologous across different species to facilitate translation of preclinical biomarkers to the clinic

Key strengths of the MASSterclass™ platform for protein quantitation:

Protein assay development of up to 50 proteins in 8 weeks
No need for antibodies
Accurate protein quantitation up to 10 pg/ml in plasma
Only 25 µl of sample volume required for quantitation of all proteins in a single assay
Analytes unambiguously identified by the mass spectrometer
Detects peptide surrogates of the protein targets (rather than the intact proteins) so less subject to variability due to sample degradation
Build assays to regions of protein sequence that are homologous across different species to facilitate translation of preclinical biomarkers to the clinic