Automated Detection of Conformational Epitopes Using Phage Display Peptide Sequences

Background: Precise determination of conformational epitopes of neutralizing antibodies represents a key step in the rational design of novel vaccines. A powerful experimental method to gain insights on the physical chemical nature of conformational epitopes is the selection of linear peptides that bind with high affinities to a monoclonal antibody of interest by phage display technology. However, the structural characterization of conformational epitopes from these mimotopes is not straightforward, and in the past the interpretation of peptide sequences from phage display experiments focused on linear sequence analysis to find a consensus sequence or common sequence motifs. Results: We present a fully automated search method, EpiSearch that predicts the possible location of conformational epitopes on the surface of an antigen. The algorithm uses peptide sequences from phage display experiments as input, and ranks all surface exposed patches according to the frequency distribution of similar residues in the peptides and in the patch. We have tested the performance of the EpiSearch algorithm for six experimental data sets of phage display experiments, the human epidermal growth factor receptor-2 (HER-2/neu), the antibody mAb Bo2C11 targeting the C 2 domain of FVIII, antibodies mAb 17b and mAb b12 of the HIV envelope protein gp120, mAb 13b5 targeting HIV-1 capsid protein and 80R of the SARS coronavirus spike protein. In all these examples th

Electrical Properties of lead free ceremics Na1−XKxNbO3, at x=0.305.

By solid state reaction method, ceramic pellets of Na0.695K0.305NbO3 have been prepared. X-ray- diffraction, Piezo properties, scanning electron microscope, and temperature dependence of dielectric constant and loss tangent of the prepared samples have been studied. It has been observed that, at the transition temperature, dielectric constant peak shifts to lower temperature, and the dielectric constant and loss tangent peak heights decrease, with  increasing frequency, and show three structural phase transitions

Dielectric properties of Na1−xKxNbO3, near x = 0.475 morphotropic phase region

11-17Sodium-potassium niobate (Na1−xKxNbO3) ceramic pellets, with x = 0.465, 0.470, 0.475, 0.480 and 0.485, were prepared by solid state reaction method with double sintering. With varying composition (x), the dielectric properties, X-ray diffraction peaks shifting pattern, in this system. Among the prepared compositions, a break in the X-ray diffraction peaks shifting tendency; minimum dielectric constant, loss tangent and dielectric conductivity indicate a morphotropic phase transition type behavior, at the composition with x = 0.475

Electrical Properties of lead free ceremics Na1−XKxNbO3, at x=0.305.

783-786By solid state reaction method, ceramic pellets of Na0.695K0.305NbO3 have been prepared. X-ray- diffraction, Piezo properties, scanning electron microscope, and temperature dependence of dielectric constant and loss tangent of the prepared samples have been studied. It has been observed that, at the transition temperature, dielectric constant peak shifts to lower temperature, and the dielectric constant and loss tangent peak heights decrease, with increasing frequency, and show three structural phase transitions

Functional classification of protein toxins as a basis for bioinformatic screening

Proteins are fundamental to life and exhibit a wide diversity of activities, some of which are toxic. Therefore, assessing whether a specific protein is safe for consumption in foods and feeds is critical. Simple BLAST searches may reveal homology to a known toxin, when in fact the protein may pose no real danger. Another challenge to answer this question is the lack of curated databases with a representative set of experimentally validated toxins. Here we have systematically analyzed over 10,000 manually curated toxin sequences using sequence clustering, network analysis, and protein domain classification. We also developed a functional sequence signature method to distinguish toxic from non-toxic proteins. The current database, combined with motif analysis, can be used by researchers and regulators in a hazard screening capacity to assess the potential of a protein to be toxic at early stages of development. Identifying key signatures of toxicity can also aid in redesigning proteins, so as to maintain their desirable functions while reducing the risk of potential health hazards

Novel neutralizing monoclonal antibodies protect rodents against lethal filovirus challenges

Filoviruses are the causative agents of lethal hemorrhagic fever in human and non-human primates (NHP). The family of Filoviridae is composed of three genera, Ebolavirus, Marburgvirus and Cuevavirus. There are currently no approved vaccines or antiviral therapeutics for the treatment of filovirus infections in humans. Passive transfer of neutralizing antibodies targeting the Ebola virus (EBOV) glycoprotein (GP) has proven effective in protecting mice, guinea pigs and NHP from lethal challenges with EBOV. In this study, we generated two neutralizing monoclonal antibodies (MAbs), termed S9 and M4 that recognize the GP of EBOV or multiple strains of Marburg virus (MARV), respectively. We characterized the putative binding site of S9 as a linear epitope on the glycan cap of the GP1 subunit of the EBOV-GP. The M4 antibody recognizes an unknown conformational epitope on MARV-GP. Additionally, we demonstrated the post-exposure protection potential of these antibodies in both the mouse and guinea pig models of filovirus infection. These data indicate that MAbs S9 and M4 would be good candidates for inclusion in an antibody cocktail for the treatment of filovirus infections