MiS-MALDI: microgel-selected detection of protein biomarkers by MALDI-ToF mass spectrometry.
Intensified efforts to decipher the origin of disease at the molecular level stimulate the emergence of more efficient proteomic technologies. To complement this, attempts are being made to identify new predictive biomarkers for building more reliable biomarker patterns. As biomarker research gathers pace an immediate interest becomes focused on platforms, which although based on mainstream approaches, are more amenable to specialist tasks. Particularly relevant this is for disease-specific biomarkers, which are present at very low concentrations in multicomponent biological fluids and require depletion protocols enabling their separation from high-abundance components. In this report, we describe a new strategy allowing the rapid detection of target protein biomarkers by MALDI-ToF mass spectrometry. The approach relies on selective sequestering of target proteins from complex media by engineered microgels, which select proteins by their size (<30 kDa) and isoelectric points (protein pI <6.5). Subsequently, biomarker-loaded microgels are subjected to direct mass-spectrometric analysis without the need for preceding protein extraction. Exemplified by a natural protein-folding motif, coiled-coil, the monitoring of hierarchical folding-dependent macromolecular systems by the approach is also shown. The described strategy offers a general rationale for versatile platforms for high throughput proteomics and holds promise for proteome fingerprinting of biomolecular interactions.
Cerasoli
Eleonora
E
National Physical Laboratory, Teddington, TW11 0LW, UK.
Rakowska
Paulina D
PD
Horgan
Adrian
A
Ravi
Jascindra
J
Bradley
Melanie
M
Vincent
Brian
B
Ryadnov
Maxim G
MG
D000595:Amino Acid Sequence; D015415:Biomarkers / Q000032:analysis; D005782:Gels / Q000737:chemistry*; D006801:Humans; D008689:Methacrylates / Q000737:chemistry*; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D015415:Biomarkers; D005782:Gels; D008689:Methacrylates; D010455:Peptides; D011506:Proteins; D011092:Polyethylene Glycols