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Proteomics

Proteomics is a branch of biotechnology that brings together the techniques of molecular biology, biochemistry and genetics to facilitate the identification and molecular characterisation of the proteins that are expressed by the genes of a particular cell under defined environmental conditions.

This data, together with downstream bioinformatic analyses, provides detailed information on protein structure and function, regulation of protein expression, post-translational modification and the molecular interactions of proteins during both normal physiological and diseased states.

The field of proteomics is dynamic and continues to evolve rapidly to address issues arising from sources ranging from genome sequence through to cell phenotype and its approaches have many potential applications in biomedical science and biotechnology.

The Moredun Proteomics Facility (MPF) was established in 2000 with financial assistance from the Scottish Government, European Regional Development Fund (ERDF) and Scottish Enterprise Edinburgh and Lothian (SEEL). The MPF provides the facilities and expertise for the investigation of infectious diseases both from pathogen and host perspectives and the facility supports and contributes to many research projects.

The MPF provides support to scientists at Moredun as well as colleagues from other research providers, universities and private commerce. The MPF welcomes collaborations with external partners and can assure complete confidentiality through signed agreements. MPF staff members are available for consultation by arrangement and can offer advice on experimental design, protocol development, data analysis, etc.

Where MPF staff members contribute towards your research, recognition through co-authorship and/or acknowledgement of MPF and its staff is required.  Click here for Guidelines. 

An overview of services available at the Moredun Proteomics Facility can be found below or contact us to discuss your specific requirements.

Services avaliable at the Moredun Proteomics Facility (MPF)

 

Pre-Fractionation of Complex Protein Mixtures

An important initial step in any proteomics workflow is pre-fractionation of complex protein mixtures, e.g. crude cell lysates, into manageable simplified samples that facilitate high resolution analysis by subsequent 2-D gel electrophoresis and/or mass spectrometry. This can be achieved in a number of ways including liquid iso-electric focusing (liquid IEF) and more traditional single and multi-dimensional chromatographic (HPLC) strategies.

  • HPLC (nano-flow; capillary-flow; microbore; analytical)
  • Liquid IEF
  • Solid Phase Extraction
  • Gel Electrophoresis

 

2-D Gel Electrophoresis

The Moredun Proteomics Facility is equipped with a number of GE Healthcare IPGPhor™ IEF (1st dimension) and MultiPhor™ and Ettan-DALT™ (2nd dimension) platforms. A wide selection of 1st and 2nd dimension gel formats is available ranging from smaller (7cm) gels for pilot experiments to large (24cm) gels for more definitive high-resolution work

  • IPGPhor™ IEF (1st dimension)
  • MultiPhor™ and Ettan-DALT™ (2nd dimension) platforms
  • Gel formats for pilot (7cm) and analytical (up to 24cm) studies

 

Liquid Chromatography

The Moredun Proteomics Facility can provide chromatographic separations on analytical, micro-bore, capillary-flow and nano-flow scales. Dionex UltiMate™ and Beckman-Coulter System-Gold HPLC platforms provide for size exclusion, ion exchange, hydrophobic interaction and reversed phase separations for a range of preparative and analytical applications. Single- and multi-dimensional chromatographic protocols can be custom-tailored to yield highly enriched target molecules.

  • Dionex UltiMate™ and BioLC
  • Beckman-Coulter System-Gold HPLC
  • Size exclusion, ion exchange, hydrophobic interaction and reversed phase separations
  • Multi-dimensional chromatographic purification strategies

 

Mass Spectrometry

The Moredun Proteomics Facility is equipped with three tandem mass spectrometers including a Bruker Ultraflex II MALDI-ToF, a Bruker maXis UHR-Q-ToF and a Bruker amaZon-ETD 3-dimensional ion trap. Collectively, these instruments enable the MPF to provide a comprehensive range of MS applications including protein identification, de novo characterisation of peptides, analysis of post-translational modifications and label-free quantitation.

  • Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-ToF MS/MS)
    • Bruker UltraflexII
  • Liquid Chromatography Electrospray ionisation MS/MS
    •  Bruker amaZon-ETD spherical ion trap
    •  Bruker maXis Ultra-High-Resolution Q-ToF

 

Applications

  • Protein identification
  • Shotgun proteome analysis
  • Label-free Quantitation
  • Protein characterisation (including Post-translational modifications)
  • T3 Protein Sequencing
  • Whole cell MALDI (microbial identification & differentiation)
  • Top-down proteomics (ETD & PTR)
  • Intact Mass Analysis
  • LC-MALDI
  • Biomarker Identification & Characterisation (ClinProt; SELDI)

 

Analytical Tools

  • MASCOT
  • ProteinScape

Publications

For a list of publications to which the MPF has contributed please click here