Recombinant Proteins Unit

>>Recombinant Proteins Unit

Recombinant proteins —those produced in the laboratory by genetic engineering using cells different from nature— have shown a high impact in basic research and in the biomedical field for drug manufacture. However, in many cases it is not possible to obtain a stable, soluble and homogeneous product, and this limits its applications. Several strategies were developed in the last decades to overcome these limitations. In this sense, our group has generated a set of vectors that facilitates cloning and allows the evaluation of several parameters that can improve the soluble expression of a target protein.

In the context of cancer therapeutic tools, our group recently focused on the generation of artificial binding proteins known as Affitins. This class of proteins has a wide range of advantages compared to classical therapeutic antibodies that could be taken into account in the development of therapeutic approaches.


Agustín Correa, PhD

Associate Technician

acorrea@pasteur.edu.uy


Claudia Ortega, PhD

Technical assistant

ckortega@pasteur.edu.uy

  • Vector Suite and Artificial Binding Proteins (ABP)
    Our laboratory has developed a set of vectors that allow the evaluation of different promoters and proteins that enhance solubility, through an efficient cloning strategy. This vector suite was extended in order to generate a set of vectors to include the evaluation of expression of recombinant proteins in different cellular compartments and cellular hosts. This helps to overcome the limitations encountered when working with a single subcellular location and a single type of host. In addition, these vectors also allow the evaluation of alternative purification strategies for the improvement of the yields of the target protein.

    In the context of cancer therapeutic tools, our group recently focused on the generation of artificial binding proteins known as Affitins. This class of proteins has a wide range of advantages compared to classical therapeutic antibodies that could be taken into account in the development of therapeutic approaches.

    Compared to classical therapeutic antibodies, Affitins are able to maintain constant high affinity even when their molecular weight remains small. This could be very useful in lymphoid neoplasms, in order to gain access to solid tissues as secondary lymphoid organs, where leukemic cells receive survival signals that acquire favourable conditions of proliferation.

    In this line, a new generation of combinatorial protein engineering technologies has recently been created in our laboratory. The results have made it possible to propose the use of these artificial binding proteins as versatile selective glycosidase inhibitors and, potentially, as enzymatic inhibitors in general, which could be foreseen for future tumour therapy strategies (Correa et al., PlosOne, 2014).

  • ÄKTAxpress / ÄKTA Pure and ÄKTA Purifier

  • Benchtop Bioreactor BIOSTAT® B plus (Prokaryotic culture)

  • CelliGen 310 Bioreactor (Eukaryotic culture)

  • BelloCell 3000 Bioreactor (Eukaryotic culture)

  • EmulsiFlex-C5 Homogenizer

  • Multitron 2 Incubated Shaker

  • Expression of recombinant proteins (PR) in prokaryotic and eukaryotic systems:

    • PR expression in E. coli
    • PR expression in mammalian cells
    • PR expression in insect cells
  • Optimization of the expression of soluble recombinant proteins

  • “Integrating IP Montevideo technologies”. Course for researchers and technicians of IP Montevideo, 2017.

  • Postgraduate course: “Introduction to the structural and functional analysis of proteins”, Institut Pasteur de Montevideo. September-November 2014.

  • Postgraduate course: “Expression, Purification and Crystallization of Recombinant Proteins by High-throughput Methodologies”. February 2013.

  • Postgraduate course: “Expression of Recombinant Proteins”. PEDECIBA-Biology- MSc in Biotechnology, Faculty of Science – Institut Pasteur de Montevideo. Nov5–Dec10, 2008.

  • 2016 – Design and development of artificial binding proteins with potential use in biomedicine. Scientific responsible: Agustín Correa. FCE_3_2016_1_125765.