PS Biomarker ServicesTM Proteomics Work Flow
Customized, Multiplexed Large Protein Analysis – Mass Spectrometry Allows Access to a Greater Proportion of the Proteome
The Proteomics work flow allows for identification and measurement of the total protein content of cells, tissues and body fluids. The classical approach to analysis of proteins is firstly to digest them into smaller fragments, and following fractionation and separation of the digests, quantitation and identification of specific peptides. The use of Mass Spectrometry permits precise and reproducible quantitation of a vast array of proteins in the proteome.
The PS Biomarker ServicesTM LC-MS/MS Proteomics work flow utilizes trypsin digestion (or other protease treatment), isobaric TMT labeling, liquid chromatography separation and enrichment, coupled with tandem mass spectrometry using the very latest and most sensitive instrumentation and protocols. For serum, plasma and CSF samples the protocol may be run with prior depletion of high abundant proteins. Click here to view the profiling and separation options, and here to view the protein identification and measurement capabilities of PS Biomarker ServicesTM.
The PS Biomarker ServicesTM GeLC-MS/MS Proteomics work flow utilizes isobaric TMT labeling at the protein level, 1-dimensional gel separation, trypsin digestion (or other protease treatment), liquid chromatography separation and enrichment, coupled with tandem mass spectrometry using the very latest and most sensitive instrumentation and protocols. For serum, plasma and CSF samples the protocol may be run with prior depletion of high abundant proteins. Click here to view the profiling and separation options, and here to view the protein identification and measurement capabilities of PS Biomarker ServicesTM.
The PS Biomarker ServicesTM 2-Dimensional Gel Proteomics work flow utilizes protein separation by isoelectric focusing followed by SDS-Polyacrylamide Gel Electrophoresis. Gels are stained with silver or fluorescent dyes and images compared to identify protein spots showing quantitative differences, cutting and trypsin digestion (or other protease treatment) of seleted spots, coupled with MALDI-TOF/TOF mass spectrometry using sensitive instrumentation and protocols. For serum, plasma and CSF samples the protocol may be run with prior depletion of high abundant proteins.
Click here to view the profiling and separation options, and
here to view the protein identification and measurement capabilities of our PS Biomarker Services
TM.
Mass Spectrometry is an extremely flexible technique that is not subject to the many interferences typical for most immunoassay platforms when analyzing complex biological sample materials. Protocols are available for a wide range of sample types including Cerebrospinal Fluid (CSF), plasma, serum, cultured cells, frozen biopsy material, urine, post mortem samples and several others.
The PS Biomarker ServicesTM Protein TMT-SRM work flow uses the high selectivity and sensitivity of Selected Reaction Monitoring (SRM) Mass Spectrometry combined with isobaric and isotopic forms of TMT labeling to deliver seamless integration of biomarker discovery and targeted assay development workflows without the need to synthesise expensive AQUA peptides. TMT-SRM methods can be developed for tens to hundreds of candidate biomarkers within a few days of completing discovery studies. After qualification a final biomarker panel can be transferred for absolute quanitative assay development by SRM or immunoassay.
Advantages of MS Proteomics Work Flow Approach:
- Does not require antibodies – relatively few proteins in the proteome have commercially available antibodies
- Access the whole proteome – mass spectrometry permits measurement of nearly all proteins and fragments
- Wide range of sample types – most samples types can easily be processed
- Unambiguous protein fragment ID & measurement – accuracy and reproducibility of MS for protein fragments much greater compared to other assay technologies
- Identify multiple fragments of the same protein – unlike immunoassays, MS can rapidly quantitate post-translationally modified variants or fragments of the same protein
- Rapid development of multiplexed assays – Using TMT-SRM we deliver shorter development time for higher content results compared to immunoassays
- No quenching interference – compared to immunoassay approach
Mass Spectrometry Proteomics Work Flow:
Applications of a Proteomics Work Flow Approach
- Target Discovery/Validation – identify and validate potential new drugable targets
- Biomarker Discovery – Mass Spectrometry allows rapid and systematic discovery of new biomarkers for specific diseases
- Drug Discovery Screening – multiplexed immunoassays are not available for all biomarkers of interest
- Clinical Diagnostic Applications – precision & reproducibility of Mass Spectrometry is ideally suited to clinical applications
- Off-target effects – monitor for non-specific and undesired drug off-targets effects of drugs
Contact us to learn more about the extensive range of Products and Services available through PS Biomarker ServicesTM or to arrange a consultation with our scientists.
