Sai H A Gandham

University of Texas Health Science Center at Houston, Houston, Texas, United States

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Publications (8)11.47 Total impact

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    ABSTRACT: Thioaptamers targeting the dengue-2 virus (DENV-2) envelope protein domain III (EDIII) were developed. EDIII, which contains epitopes for binding neutralizing antibodies, is the putative host-receptor binding domain and is thus an attractive target for development of vaccines, anti-viral therapeutic and diagnostic agents. Thioaptamer DENTA-1 bound to DENV-2 EDIII adjacent to a known neutralizing antibody binding site with a dissociation constant of 154 nM.
    Biochemical and Biophysical Research Communications 09/2014; · 2.28 Impact Factor
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    Muniasamy Neerathilingam, Sumukh Mysore, Sai Hari A. Gandham
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    ABSTRACT: Inclusion bodies (IBs) are commonly formed in Escherichiacoli due to over expression of recombinant proteins in non-native state. Isolation, denaturation and refolding of these IBs is generally performed to obtain functional protein. However, during this process IBs tend to form non-specific interactions with sheared nucleic acids from the genome, thus getting carried over into downstream processes. This may hinder the refolding of IBs into their native state. To circumvent this, we demonstrate a methodology termed soni-removal which involves disruption of nucleic acid–inclusion body interaction using sonication; followed by solvent based separation. As opposed to conventional techniques that use enzymes and column-based separations, soni-removal is a cost effective alternative for complete elimination of buried and/or strongly bound short nucleic acid contaminants from IBs.
    Biochemical and Biophysical Research Communications 05/2014; 448(1):45–49. · 2.28 Impact Factor
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    ABSTRACT: Inclusion bodies (IBs) are commonly formed in E.coli during over expression of recombinant proteins in non-native state. Isolation, denaturation and refolding of these IBs is generally practiced to obtain functional protein. However, along this process IBs tend to form non-specific interactions with sheared nucleic acids from the genome, thus getting carried over into down stream processes. This may hinder the refolding of IBs into their native state. To circumvent this, we demonstrate a methodology termed soni-removal which involves disruption of nucleic acid-inclusion body interaction using sonication; followed by solvent based separation. As apposed to conventional techniques that use enzymes and column-based separations, soni-removal is a cost effective alternative for complete elimination of buried and/or strongly bound short nucleic acid contaminants from IBs.
    Biochemical and Biophysical Research Communications 04/2014; · 2.28 Impact Factor
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    ABSTRACT: NF-κB plays a crucial role in regulating various biological processes including innate and adaptive immunity, inflammation, stress responses, B-cell development, and lymphoid organogenesis. Currently, several assays like electrophoretic mobility shift assay (EMSA), enzyme-linked immunosorbent assay (ELISA), fluorescence resonance energy transfer (FRET) and time-resolved fluorescence resonance energy transfer (TR-FRET) are widely used for studying the NFκB intraction with β-IFN-κB binding oligo. Each of these techniques have varying utility with distinct strengths and weaknesses. We describe a method AlphaLISA to identify NFκB p50 protein and β-IFN-κB binding oligo sequence and interaction is efficient at a given concentration (10 nM) in the EMSA and Biacore’s SPR assays. The method has many advantages such as use of small volume, high throughput (HTP), convenience of sample preparation and data analysis.
    Journal of Analytical Sciences, Methods and Instrumentation 09/2013; 3(3):173-178.
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    ABSTRACT: The structure of recombinant domain III of the envelope protein (rED3) of yellow fever virus (YFV), containing the major neutralization site, was determined using NMR spectroscopy. The amino acid sequence and structure of the YFV-rED3 shows differences from ED3s of other mosquito-borne flaviviruses; in particular, the partially surface-exposed BC loop where methionine-304 and valine-324 were identified as being critical for the structure of the loop. Variations in the structure and surface chemistry of ED3 between flaviviruses affect neutralization sites and may affect host cell receptor interactions and play a role in the observed variations in viral pathogenesis and tissue tropism.
    Virology 10/2009; 394(1):12-8. · 3.35 Impact Factor
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    ABSTRACT: Nearly complete backbone and side chain resonance assignments have been obtained for the third domain, residues M289-K400, of the envelope protein from the sylvatic strain (P72-1244) of the dengue 1 virus, containing mutations N336S and E370K, using double- and triple-resonance spectroscopy.
    Biomolecular NMR Assignments 01/2009; 2(2):155-7. · 0.64 Impact Factor
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    ABSTRACT: Nearly complete backbone and sidechain resonance assignments have been obtained for the third domain, residues S288-K398, of the envelope protein from the Asibi strain of yellow fever virus using double- and triple-resonance spectroscopy.
    Biomolecular NMR Assignments 08/2007; 1(1):49-50. · 0.64 Impact Factor