Publications on the project |
084 Isolation and characterisation of recombinant single chain variable fragment antibodies (scFv) against human heparin-binding EGF-like growth factor (HB-EGF) |
Authors: | O.S. Oliinyk, A.A. Kaberniuk, D.V. Kolibo, S.V. Komisarenko | |
Summary: | Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the EGF family of growth factors, wich plays an important role in many physiological and pathological processes. Antibodies are an important tool for investigation of HB-EGF functions at normal and pathological processes. The aim of this study was to obtain and to study some properties of recombinant single-chain variable fragment (scFv) antibodies specific to HB-EGF. For this purpose a phage-displayed immune library of mice immunized with recombinant human soluble HB-EGF was generated. The library was screened against HB-EGF, and 2 groups of antibody clones with different fingerprinting patterns were isolated. Two clones – one from each group (scFv 5A and scFv 7E) were analyzed. It was shown that these scFv antibodies could bind both – human and mouse HB-EGF. Both clones recognized target antigen in Western blot, but only under non-reducing conditions. Specific binding of scFv 5A and scFv 7E to Vero cells with high level of HB-EGF on their surface was also demonstrated by confocal microscopy.
In this study scFv antibodies against soluble HB-EGF were obtained. They were suitable for ELISA, Western blot and immunocytochemistry applications. These scFv could be further used as a tool in HB-EGF research and, possibly, for the development of new diagnostic and therapeutic agents. | |
Keywords: | Heparin-binding epidermal growth factor-like growth factor, scFv-antibodies, phage display | |
Edition: | Biotechnologia Acta | | | 2012,
Т.5, №6. – С. 47-56.,Ukrainian |
084 Obtaining and characterization of recombinant fluorescent derivatives of soluble human HB-EGF |
Authors: | N. V. Korotkevich, A. Ju. Labyntsev, D. V. Kolibo, S. V. Komisarenko | |
Summary: | Heparin-binding EGF-like growth factor (HB-EGF) belongs to the epidermal growth factor receptor family and is synthesized as a transmembrane precursor proHB-EGF. Binding of sHB-EGF with receptors EGFR and HER-4 results in formation of ligand-receptor complexes and activation of signaling pathways and thereby promotes survival, proliferation and migration of cells. The aim of the study was to obtain the recombinant fluorescent derivatives of full-length soluble sHB-EGF – mCherry-sHB-EGF and truncated sHB-EGFΔ84-106 (without heparin-binding domain) – mCherry-sHB-EGFΔ84-106. | |
Keywords: | heparin-binding EGF-like growth factor (HB-EGF), epidermal growth factor receptor | |
Edition: | Biotechnologia Acta | | | 2014,
Т. 7, № 2, С. 46-53,Ukrainian |
084 Recombinant antibodies for fundamental and applied research |
Authors: | Oliinyk O.S. | |
Summary: | Recombinant antibody technology is a powerful approach for generation new diagnostic and therapeutic reagents. Recombinant antibodies could be applied in the construction of drug delivery systems, immunotoxins, as intrabodies. They also could be genetically fused to the different marker molecules, such as fluorescent proteins or enzymes for one-step immunodetection of antigens. This paper provides an overview of the properties and current studies on the application of recombinant antibodies. | |
Keywords: | recombinant antibodies, scFv, immunoliposomes, immunotoxins, intrabodies | |
Edition: | Bulletin of the National Academy of Sciences of Ukraine | | | 2014,
№9. – С 77-88.,Ukrainian |
084 Immunoliposomes for the targeted delivery of bioactive compounds to tumor cells |
Authors: | Oliinyk O.S., Labyntsev A.J., Manoilov K.Y., Kolibo D.V., Komisarenko S.V. | |
Summary: | Increased expression of Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is observed in a number of tumor types. Thus, this molecule could be an attractive target for antibody-based drug delivery into tumor cells. In this study we generated single-chain variable fragment antibodies (scFv) to HB-EGF and used them for the construction of immunoliposomes.
These immunoliposomes were evaluated for their specific binding and internalization for HB-EGF-expressing Vero cells. By means of flow cytometry and fluorescence microscopy it was demonstrated that HB-EGF-targeted immunoliposomes in contrast to nontargeted liposomes have been efficiently bound and internalized by Vero cells. These results indicate that HB-EGF-targeted immunoliposomes provide efficient targeted delivery of anticancer drugs into HB-EGF-expressing cells. | |
Keywords: | immunoliposome, targeted delivery, scFv-antibody, HB-EGF | |
Edition: | “Nanoscale systems and nanomaterials: research in Ukraine systems and nanomaterials: research in Ukraine” (ed. by A.G. Naumovets): NAS of Ukraine. – Kyiv, “Academperiodika” | | | 2014,
510-514,Russian |
The events in the framework of the project |
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084 5. Nanobiotechnology Purpose: Expected results:Issue of new types of products: methods, theories Stage 1: Stage 2: Stage 3: Stage 4: Stage 5:
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