Bioengineering Department. Operations

As implied by the type of research, the Bioengineering Department is divided into the following laboratories:

• Immunology Laboratory
• Tissue Culture Laboratory
• Genetic Engineering Laboratory
• Protein Research Laboratory
• Microbiology Laboratory
• Biochemistry Laboratory
• Bacterial Genetics Laboratory
• Analytical Laboratory 1
• Mass Spectrometry Laboratory

Immunology Laboratory

The laboratory deals with:

1. Preparation of the antibodies for analytical purposes (control of the immunization efficacy in the procedure of obtaining polyclonal sera, purification of the polyclonal and monoclonal antibodies using affinity chromatography method, antibody labelling).
2. Development of ELISA tests for purity evaluation of recombined pharmaceuticals (determining the level of contamination with proteins of Escherichia coli host strain (ECP), determination of the remnants of enzymatic preparations used in the technology) and other proteins.
3. Determination of the level of recombined protein expression in the transgenic material by immunological techniques.
4. Evaluation of the immunogenicity of recombined antigens by parenteral immunization.

The laboratory provides services within the operations described in item 1 and 2.

Tissue Culture Laboratory

The laboratory deals with:

1. Development of determination methods for protein drug biological activity of cultured cells.
2. Production of relevant monoclonal antibodies necessary for analyses.
3. Undertaking attempts to obtain stable expressions of the selected proteins in the cultured cells.

Genetic Engineering Laboratory

The laboratory deals with:

1. Isolation of plasmid and chromosomal DNA.
2. Construction of production strains of E. coli producing proteins and peptides useful in biotechnology, medicine and science.
   
3. Development of new expression vectors.
4. Searching for the specific proteases useful in e.g. production of the medications (e.g. obtaining production strains of E. coli synthesizing deubiquitinating protease that is an analogue of the yeast protease UBP1).
5. DNA sequencing

The laboratory provides services within the operations described in item 2.

Protein Laboratory

The laboratory deals with:

1. Development, in laboratory scale, of the method of isolation and purification of proteins and peptides produced by production strains and with consideration of conditions necessary for the transfer to industrial scale using various chromatographic techniques: ion exchange, hydrophobic, molecular filtering, affinity and other, conducted under low-, medium- and high-pressure conditions (FPLC and HPLC) at room temperature or at +4°C.

The laboratory provides services within the operations described in item 1.

Microbiology Laboratory

The laboratory deals with:

1. Maintenance of vitality of the IBA microorganisms collection (over 800 strains, including 145 under reference IAW in the Catalogue of Polish Collection of Microorganisms since 1990, membership in ECCO since 1995, membership in WFCC since 1996).
2. Preparation of strains for sale and exchange within ECCO and WFCC, sharing the strains with other scientific and research establishments, conducting contracted research.

The laboratory provides services within the operations described in item 2.

Biochemistry Laboratory

The laboratory deals with:

1. Genetic modification within the vector leading to optimization of the expression level of created protein, as well as improvement in the production strain stability.
2. Development of the new expression vectors.
3. Protein hyperexpression in prokaryotic systems.
4. Molecular analysis of antigen suitability for construction of new generation vaccines (subunit vaccines) and the development of new carriers to deliver vaccine antigens through mucosal membranes.
5. Obtaining proteins with significant therapeutic action.

Bacterial Genetics Laboratory

The laboratory deals with:

1. Analysis of the factors determining drug resistance of the Enterobacteriaceae clinical strains (plasmids, integrons).
2. Analysis of plasmids isolated from natural environments (isolation, sequencing, property analysis).
3. Development of in-house expression systems with the use of natural plasmids.
4. Evaluation of the replication biology of cryptic plasmids.

Analytical Laboratory 1

The laboratory deals with:

1. Construction of E. coli production strains producing proteins and peptides useful in biotechnology, medicine and science.
2. Quantitative determinations of plasmid and bacterial DNA contaminations in biosimilar pharmaceuticals by the quantitative PCR method.

The laboratory provides services within the operations described in item 1 and 2.

Mass Spectrometry Laboratory

The laboratory deals with:

1. Evaluation of proteins and peptides within the scope of:
   • separation,
   • identification,
   • quantitative analysis,
   • sequence analysis,
   • modification studies.
2. Determination of the molecular mass of peptides, proteins, nucleotides, oligosaccharides and lipids.
3. Determination of the mass profiles of proteins and peptide maps.
4. SDS analysis (determination of the molecular mass of evaluated protein, determination of its the percentage of contamination).
5. Isoelectric focusing (determination of the isoelectric point of evaluated protein and determination of potential isoforms).
6. Preliminary analysis of protein profiles (determination of additional derivative forms e.g. deamidated, cut off).
7. Analysis of antibody purity and heterogeneity.
8. 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).