The Experts below are selected from a list of 134022 Experts worldwide ranked by ideXlab platform

Vladimir Kery - One of the best experts on this subject based on the ideXlab platform.

  • Automated 96-Well Purification of Hexahistidine-Tagged Recombinant Proteins on MagneHis Ni 2 +-Particles
    Methods in molecular biology (Clifton N.J.), 2009
    Co-Authors: Chiann-tso Lin, Priscilla A. Moore, Vladimir Kery
    Abstract:

    Functional genomics and the application of high-throughput (HT) approaches to solve biological and medical questions are the main drivers behind the increasing need for HT parallel expression and purification of Recombinant Proteins. Automation is necessary to facilitate this complex multistep process. We describe, in detail, an HT-automated purification of hexahistidine-tagged Recombinant Proteins using MagneHis Ni-Particles and the Biomek FX robot. This procedure is universally applicable to hexa-histidine-tagged Recombinant Proteins with the tag positioned at either the N- or C-terminus. With minor modifications, the automated protein purification protocol presented in this chapter could be adapted to purify Recombinant Proteins bearing other tags than hexahistidine and/or other expression systems than E. coli.

  • automated 96 well purification of hexahistidine tagged Recombinant Proteins on magnehis ni 2 particles
    Methods of Molecular Biology, 2009
    Co-Authors: Chiann-tso Lin, Priscilla A. Moore, Vladimir Kery
    Abstract:

    Functional genomics and the application of high-throughput (HT) approaches to solve biological and medical questions are the main drivers behind the increasing need for HT parallel expression and purification of Recombinant Proteins. Automation is necessary to facilitate this complex multistep process. We describe, in detail, an HT-automated purification of hexahistidine-tagged Recombinant Proteins using MagneHis Ni-Particles and the Biomek FX robot. This procedure is universally applicable to hexa-histidine-tagged Recombinant Proteins with the tag positioned at either the N- or C-terminus. With minor modifications, the automated protein purification protocol presented in this chapter could be adapted to purify Recombinant Proteins bearing other tags than hexahistidine and/or other expression systems than E. coli.

A. I. Kornelyuk - One of the best experts on this subject based on the ideXlab platform.

  • Stabilization of AIMP1/p43 and EMAP II Recombinant Proteins in the complexes with polysaccharide dextran-70.
    Pharmacological reports : PR, 2020
    Co-Authors: L. A. Kolomiiets, N. V. Vorobyova, D. M. Lozhko, V. M. Zayets, A. I. Kornelyuk
    Abstract:

    BACKGROUND Protein-based pharmaceuticals are among the fastest growing categories of therapeutic agents in the clinic and as commercial products, and typically target high-impact areas such as various cancers, autoimmune diseases and metabolic disorders. The aim of our work was to explore the possibility of reducing the level of aggregation and improve the stability of the Recombinant Proteins AIMP1/p43 (aminoacyl-tRNA synthetase complex component of the higher eukaryotes) and antitumor cytokine EMAP II (proteolytic cleavage product of AIMP1/p43) in combination with dextran-70 polysaccharide for structural-functional research and development of new sustainable biomedical products. METHODS We studied interaction strength between these Recombinant Proteins with polymer by fluorescence spectroscopy and molecular docking. RESULTS During experimental studies, optimal concentration ratio of AIMP1/p43 and EMAP II Recombinant Proteins with dextran-70 in which Proteins bind to ligand and form complex was established. As a result of molecular docking investigations, spatial structure of the AIMP1/p43-dextran-70 and EMAP II-dextran-70 complexes was obtained and their binding energy was evaluation. CONCLUSIONS The effect of temperature increase on the stability of these two complexes was determined by fluorescence spectroscopy method. It was found that dextran-70 specifically connects with Recombinant Proteins. Binding stoichiometry of dextran-70 with protein is about 1:1, which confirms the formation of a specific complex.

  • stabilization of aimp1 p43 and emap ii Recombinant Proteins in the complexes with polysaccharide dextran 70
    Pharmacological Reports, 2020
    Co-Authors: L. A. Kolomiiets, N. V. Vorobyova, D. M. Lozhko, V. M. Zayets, A. I. Kornelyuk
    Abstract:

    BACKGROUND Protein-based pharmaceuticals are among the fastest growing categories of therapeutic agents in the clinic and as commercial products, and typically target high-impact areas such as various cancers, autoimmune diseases and metabolic disorders. The aim of our work was to explore the possibility of reducing the level of aggregation and improve the stability of the Recombinant Proteins AIMP1/p43 (aminoacyl-tRNA synthetase complex component of the higher eukaryotes) and antitumor cytokine EMAP II (proteolytic cleavage product of AIMP1/p43) in combination with dextran-70 polysaccharide for structural-functional research and development of new sustainable biomedical products. METHODS We studied interaction strength between these Recombinant Proteins with polymer by fluorescence spectroscopy and molecular docking. RESULTS During experimental studies, optimal concentration ratio of AIMP1/p43 and EMAP II Recombinant Proteins with dextran-70 in which Proteins bind to ligand and form complex was established. As a result of molecular docking investigations, spatial structure of the AIMP1/p43-dextran-70 and EMAP II-dextran-70 complexes was obtained and their binding energy was evaluation. CONCLUSIONS The effect of temperature increase on the stability of these two complexes was determined by fluorescence spectroscopy method. It was found that dextran-70 specifically connects with Recombinant Proteins. Binding stoichiometry of dextran-70 with protein is about 1:1, which confirms the formation of a specific complex.

Mingming Dong - One of the best experts on this subject based on the ideXlab platform.

  • A chemoenzymatic approach enables the site‐specific conjugation of Recombinant Proteins
    Electrophoresis, 2019
    Co-Authors: Yan Wang, Mingming Dong, Mingliang Ye
    Abstract:

    Many biotechniques including protein microarray, drug screening, biosensors rely on the immobilization of Recombinant Proteins on the solid supports. It is well known that random orientation of the immobilized Proteins could impair their biologic functions. Thus, it is very important to develop new site-specific immobilization approach. In this study, we presented a chemoenzymatic approach for site-specific conjugation of Recombinant Proteins onto solid support. In this strategy, the affinity tag on Recombinant protein was enzymatically cleaved to expose the N-terminal serine, which was oxidized to carry an aldehyde group and was then covalently coupled to hydrazide resin through hydrazone ligation. As this approach takes advantage of the most frequently used TEV protease, it requires no further sequence design on Recombinant protein. This method was validated by site specific coupling of a synthetic peptide and a Recombinant protein onto solid supports. It was found that the site specific immobilized SH2 domain is functional and could be used to enrich tyrosine phosphorylated peptides.

  • A chemoenzymatic approach enables the site-specific conjugation of Recombinant Proteins.
    Electrophoresis, 2019
    Co-Authors: Yan Wang, Mingming Dong
    Abstract:

    Many biotechniques including protein microarray, drug screening, biosensors rely on the immobilization of Recombinant Proteins on the solid supports. It is well known that random orientation of the immobilized Proteins could impair their biologic functions. Thus, it is very important to develop new site-specific immobilization approach. In this study, we presented a chemoenzymatic approach for site-specific conjugation of Recombinant Proteins onto solid support. In this strategy, the affinity tag on Recombinant protein was enzymatically cleaved to expose the N-terminal serine, which was oxidized to carry an aldehyde group and was then covalently coupled to hydrazide resin through hydrazone ligation. As this approach takes advantage of the most frequently used TEV protease, it requires no further sequence design on Recombinant protein. This method was validated by site specific coupling of a synthetic peptide and a Recombinant protein onto solid supports. It was found that the site specific immobilized SH2 domain is functional and could be used to enrich tyrosine phosphorylated peptides.

Chiann-tso Lin - One of the best experts on this subject based on the ideXlab platform.

  • Automated 96-Well Purification of Hexahistidine-Tagged Recombinant Proteins on MagneHis Ni 2 +-Particles
    Methods in molecular biology (Clifton N.J.), 2009
    Co-Authors: Chiann-tso Lin, Priscilla A. Moore, Vladimir Kery
    Abstract:

    Functional genomics and the application of high-throughput (HT) approaches to solve biological and medical questions are the main drivers behind the increasing need for HT parallel expression and purification of Recombinant Proteins. Automation is necessary to facilitate this complex multistep process. We describe, in detail, an HT-automated purification of hexahistidine-tagged Recombinant Proteins using MagneHis Ni-Particles and the Biomek FX robot. This procedure is universally applicable to hexa-histidine-tagged Recombinant Proteins with the tag positioned at either the N- or C-terminus. With minor modifications, the automated protein purification protocol presented in this chapter could be adapted to purify Recombinant Proteins bearing other tags than hexahistidine and/or other expression systems than E. coli.

  • automated 96 well purification of hexahistidine tagged Recombinant Proteins on magnehis ni 2 particles
    Methods of Molecular Biology, 2009
    Co-Authors: Chiann-tso Lin, Priscilla A. Moore, Vladimir Kery
    Abstract:

    Functional genomics and the application of high-throughput (HT) approaches to solve biological and medical questions are the main drivers behind the increasing need for HT parallel expression and purification of Recombinant Proteins. Automation is necessary to facilitate this complex multistep process. We describe, in detail, an HT-automated purification of hexahistidine-tagged Recombinant Proteins using MagneHis Ni-Particles and the Biomek FX robot. This procedure is universally applicable to hexa-histidine-tagged Recombinant Proteins with the tag positioned at either the N- or C-terminus. With minor modifications, the automated protein purification protocol presented in this chapter could be adapted to purify Recombinant Proteins bearing other tags than hexahistidine and/or other expression systems than E. coli.

L. A. Kolomiiets - One of the best experts on this subject based on the ideXlab platform.

  • Stabilization of AIMP1/p43 and EMAP II Recombinant Proteins in the complexes with polysaccharide dextran-70.
    Pharmacological reports : PR, 2020
    Co-Authors: L. A. Kolomiiets, N. V. Vorobyova, D. M. Lozhko, V. M. Zayets, A. I. Kornelyuk
    Abstract:

    BACKGROUND Protein-based pharmaceuticals are among the fastest growing categories of therapeutic agents in the clinic and as commercial products, and typically target high-impact areas such as various cancers, autoimmune diseases and metabolic disorders. The aim of our work was to explore the possibility of reducing the level of aggregation and improve the stability of the Recombinant Proteins AIMP1/p43 (aminoacyl-tRNA synthetase complex component of the higher eukaryotes) and antitumor cytokine EMAP II (proteolytic cleavage product of AIMP1/p43) in combination with dextran-70 polysaccharide for structural-functional research and development of new sustainable biomedical products. METHODS We studied interaction strength between these Recombinant Proteins with polymer by fluorescence spectroscopy and molecular docking. RESULTS During experimental studies, optimal concentration ratio of AIMP1/p43 and EMAP II Recombinant Proteins with dextran-70 in which Proteins bind to ligand and form complex was established. As a result of molecular docking investigations, spatial structure of the AIMP1/p43-dextran-70 and EMAP II-dextran-70 complexes was obtained and their binding energy was evaluation. CONCLUSIONS The effect of temperature increase on the stability of these two complexes was determined by fluorescence spectroscopy method. It was found that dextran-70 specifically connects with Recombinant Proteins. Binding stoichiometry of dextran-70 with protein is about 1:1, which confirms the formation of a specific complex.

  • stabilization of aimp1 p43 and emap ii Recombinant Proteins in the complexes with polysaccharide dextran 70
    Pharmacological Reports, 2020
    Co-Authors: L. A. Kolomiiets, N. V. Vorobyova, D. M. Lozhko, V. M. Zayets, A. I. Kornelyuk
    Abstract:

    BACKGROUND Protein-based pharmaceuticals are among the fastest growing categories of therapeutic agents in the clinic and as commercial products, and typically target high-impact areas such as various cancers, autoimmune diseases and metabolic disorders. The aim of our work was to explore the possibility of reducing the level of aggregation and improve the stability of the Recombinant Proteins AIMP1/p43 (aminoacyl-tRNA synthetase complex component of the higher eukaryotes) and antitumor cytokine EMAP II (proteolytic cleavage product of AIMP1/p43) in combination with dextran-70 polysaccharide for structural-functional research and development of new sustainable biomedical products. METHODS We studied interaction strength between these Recombinant Proteins with polymer by fluorescence spectroscopy and molecular docking. RESULTS During experimental studies, optimal concentration ratio of AIMP1/p43 and EMAP II Recombinant Proteins with dextran-70 in which Proteins bind to ligand and form complex was established. As a result of molecular docking investigations, spatial structure of the AIMP1/p43-dextran-70 and EMAP II-dextran-70 complexes was obtained and their binding energy was evaluation. CONCLUSIONS The effect of temperature increase on the stability of these two complexes was determined by fluorescence spectroscopy method. It was found that dextran-70 specifically connects with Recombinant Proteins. Binding stoichiometry of dextran-70 with protein is about 1:1, which confirms the formation of a specific complex.