Proteomics
Supervisor

Proteomics

The structural analysis of peptides and proteins, and the identification of unknown proteins are among the most important aspects of biotechnology for both basic and applied research. The Proteomic and Mass Spectrometry Facility at CEINGE provides a series of high technological analytical services addressed to the structural characterization of peptides, proteins and glycoproteins based on the integration of classical biochemical procedures with advanced mass spectrometric methodologies. These analytical services are grouped in the following categories:

Mass Spectrometry Services;

Proteomic Services.

 

All analytical services include:

Appropriate scientific consultance for customers; Interpretation of the results; Storage of data for at least three years; Data conversion to publication quality forms. A scientific report on the analytical data signed by the Responsible of the Facility is available on request.

Special agreement can be stipulated for large numbers of samples.

When complex Services not precisely fitting the standard list are needed, the most suitable strategies to approach the problem will be identified and individual quotations will be provided according to the specific needs.

 

Mass Spectrometry Services

 

Accurate Molecular Weight determination

Determination of the accurate molecular mass of peptides and proteins by either Electrospray or MALDI mass spectrometric analyses. Desalting of the samples by HPLC is also included.

 

Peptide mapping, post-translational modifications and variants.

Complete characterisation of native and recombinant proteins including analysis of protein variants and mutants, identification of post-translational modifications and proteolytic processing, and assignemnt of S-S bridges by MALDI mapping. Pre-treatment of the sample either in solution or in gel and enzymatic hydrolysis are also included.

 

Peptide sequencing by tandem MS

Direct analysis of purified samples by tandem mass spectrometry experiments using either MALDI-TOF/TOF or ES-MS/MS techniques. When needed, HPLC separation of peptides is also performed.

 

Analysis of Glycoproteins.

Complete characterisation of the oligosaccharide moieties in glycoproteins. The Service includes: Protein digestion, enzymatic stripping of glycoforms, MALDI/MS profiling of glycoforms, structural determination of individual glycoforms by MALDI-TOF/TOF techniques.

 

Sugar analysis.

Determination of the monosaccharide composition of glycoproteins by chemical hydrolysis, sugar derivatization and GC-MS analysis.

 

Analysis of metabolytes.

Pre-treatment of the sample, chemical derivatization and GC-MS analysis  or direct investigation by LC-MS/MS techniques in MRM scan mode. Quantitative determination of metabolytes can also be performed on request.

 

Proteomic Services

Identification of proteins by LCMS/MS.

Identification of proteins either in solution opr following fractionation by 1D or 2D electrophoretic methodologies. Excission of protein band(s), in situ digestion, LC-MS/MS analysis of the peptide mixtures, Database Search by Mascot software.

 

Preparation of protein samples for identification by SDS-PAGE.

Separation of protein samples by SDS-PAGE for identification of protein band(s)

 

Quantitative Differential Proteomic Analyses.

Identification and quantification of proteins differentially expressed in cell lines or tissues by tandem mass spectrometry methodologies according to the label free approach. The entire protein contents is extracted from samples, purified by solid phase extraction, digested with trypsin and subjected to LC-MS/MS analysis. Label free quantification of the differentially expressed proteins is performed by evaluating the current intensity associated with each peptide according to the extracted ionic method (XIC) or the number of fragmentation observed for each peptide ion (spectral counts, SpC).