Products and Services. Genetic Engineering

The Bioengineering Department has the capacity and expertise to provide the following services:

• Construction of production strains of Escherichia coli producing specific proteins or peptides based on in-house IBA patented expression vectors or other;
• Control of the production strain stability;
• Periodic control of correctness of the protein encoding sequence in expression vector as required for drugs by Good Manufacturing Practice (GMP);
• Development of analytical methods for an individual protein or peptide drug, to include quantitative evaluation of the host protein contamination level, and plasmid and genome DNA contamination levels as required by the European Medicines Agency, and determinations or development of the ELISA test and the production of monoclonal antibodies used in this test;
• Examination of biological activity of protein drugs manufactured on cultured cells;
• Development of ELISA immunological tests intended for determination of other contaminations in evaluated drugs, for example the content of enzymes used during production;
• Development of purification methods for specific proteins produced by the production strains of E. coli, to include limitations related to the transfer of a process from laboratory scale to industrial scale;

• Assessment of proteins and peptides in terms of:
  • Separation/purification by the methods based on protein or peptide properties (size, solubility, affinity and charge),
  • Identifications in the range from 600 Da to 200 000 Da and with accuracy up to 1000 ppm for a linear detector (masses up to 200 kDa) and 50 ppm for a reflector detector (masses up to 8000 Da),
  • Quantitative analysis by dedicated markers,
  • Analysis of amino acid sequence and modification evaluation by ProteinPilot™Software, MASCOT® and MASCOTDistiller®,
• Determination of peptide and protein molecular mass (M < 200 000 Da), oligonucleotides, oligosaccharides, polar polymers containing heteroatom such as polycarbonates, polyolefin oxides and cycloolefins (M < 20 000 Da), calixarenes and with lower efficacy, determination of lipids, polyacids, polyesters, low-polar polymers and metallo-organic compounds;
• Determination of protein mass profiles and peptide maps with the use of modern devices for the fractioning of large amounts of proteins (about 5 mL with concentration of up to 1 mg/mL) in respect of isoelectric point and hydrophobicity;
• SDS analysis (determination of the molecular mass of evaluated protein, determination of the percentage of contaminations);
• Isoelectric focusing (determination of isoelectric point of evaluated protein and determination of potential isoforms);
• Preliminary analysis of protein profiles (determination of additional derivative forms e.g. deamidated, cut off);
• Analysis of antibody purity and heterogeneity;
  
• DNA analysis:

• fragments of single-strand DNA by UV detection
• fragments of double-strand DNA by UV detection
• fragments of double-strand DNA by laser-induced fluorescence (LIF).

For any of these services please contact:

Head of the Bioengineering Department
Prof. Grażyna Płucienniczak
phone: +48 (22) 37 86 220
mail: plucienniczakg@iba.waw.pl