Analytical Services

Due to its importance in scientific research and in the quality control of synthetic peptides and/or recombinant proteins, the structural analysis of peptides and proteins constitutes one of the most relevant topics in biotechnology. We offer a wide range of advanced technical services which are very important for the structural characterization and development of bio-molecules, such as peptides and proteins. These services are based on the integration of classical biochemical techniques, such as N-terminal protein sequencing, with advanced mass spectrometry procedures.

The sequence of the first 10-20 residues at the N-terminal of a protein gives sufficient information to allow the following:

  • assigning the site of proteolytic maturation
  • synthesizing oligonucleotide probes for specific cDNA isolation
  • identifying the protein in databases
  • aligning the protein against DNA sequences
  • assessing the purity of the peptide/protein

Additionally, the determination of the accurate molecular weight of a peptide or a protein gives the following analytical information:

  • definition of the purity of the peptide/protein
  • quality control for peptide synthesis
  • confirmation of the protein sequence
  • control of the structure of recombinant proteins
  • characterization of genetic variants or mutant proteins in which there is an amino acid substitution or modification
  • definition of the proteolytic processing of the protein, both at the N- and C-termini
  • identification of post-translational maturation events or chemically induced changes
  • identification of the glycosylation pattern of glycoproteins.

N-terminal Sequencing

The N-terminal sequence of peptides and/or proteins (5-20 residues) can be carried out on samples purified by HPLC or 1D and 2D SDS-PAGE, followed by electroblotting on PVDF (Polyvinylidene difluoride) membranes using the well known EDMAN degradation method. Pre-treatment of the sample, i.e. reduction and alkylation of the cysteine residues, is also performed.
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Molecular Weight Determination

The accurate molecular weight of peptides and proteins is determined by electrospray (ESMS) and MALDI-TOF mass spectrometry analysis of crude or HPLC purified samples. Pre-treatment of the samples and/or reverse-phase HPLC purification can also be performed. Identification of modified components or contaminating species is also provided.
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Protein Identification

Proteins purified by HPLC or excised from a mono or bi-dimensional electrophoresis gel can be digested in vitro by proteolytic enzymes, such as trypsin, yielding a mixture of peptides generated on the basis of the presence of Arginine and Lysine residues in the original protein sequence. The peptide mixture is then analysed by MALDI-TOF mass spectrometry which gives the accurate molecular weight of some or most of the peptides (mass spectrometry fingerprinting). The comparison of the experimental molecular weights with those generated by virtual computer theoretical trypsin digestions of the protein sequences present in public databases, allows us to determine the nature of the protein sample and to precisely identify the protein target.
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Glycoprotein Analysis

The definition of the entire structure of the oligosaccharide moieties released from glycoproteins is achieved by determining the microheterogeneity of the different structures, the composition of the monosaccharides present, the branching pattern, the sequence of the antennae, and the possible occurrence of modifying groups, such as sulphate, phosphate, acetyl groups etc. Furthermore, the linkage positions of the individual sugars in the oligosaccharide chains and the glycosylation sites in the protein sequence are also determined.
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Analysis of proteins – peptides - sugars
Cat # Description Prices €
PRS-2201 Setup for protein/peptide sequencing 120,00
PRS-2202 N-terminal sequencing of protein/peptides on PVDF (per aa) 80,00
PRS-2203 SDS-PAGE separation of proteins and protein transfer on PVDF membranes 500,00
PRS-2204 Sample processing for internal sequencing (enzymatic hydrolysis, HPLC separation of peptides mixture, MALDI-ToF analysis of each fraction) 600,00
PRS-2205 Peptide sequencing by tandem MS (peptide separation by HPLC, fragmentation experiments by MS/MS techniques) 450,00
PRS-2206 Mass spectrometry analysis of peptides (sample purification and MALDI-ToF) 100,00
PRS-2207 Mass spectrometry analysis of proteins (sample purification and ESMS analysis) 180,00
PRS-2208 Complete peptide mapping (pre-treatment either in solution or in gel, single/double enzymatic hydrolysis, MALDI-ToF mapping) 1.400,00
PRS-2209 Identification of protein by MALDI-ToF mapping (protein digestion, MALDI-ToF analysis of peptides mixture, database search) 850,00
PRS-2210 Identification of protein (>=10 spots) by MALDI-ToF mapping (protein digestion, MALDI-ToF analysis of peptides mixture, database search) 550,00
PRS-2211 Identification of proteins by LCMS/MS (protein digestion, MALDI-ToF analysis of peptides mixture, LCMS/MS analysis, database search) 1.300,00
PRS-2212 Sugar analysis (Determination of the sugar composition in glycoproteins by chemical hydrolysis, derivatization and GCMS analysis) 600,00
PRS-2213 Analysis of glycoproteins (protein digestion, enzymatic stripping of glycoforms, MALDI-MS analysis of glycoforms) 600,00
PRS-2214 GCMS analysis (sample pre-treatment, including derivatization and GCMS analysis) 500,00

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