Day 1 :
Washington State University, USA
Keynote: Approach Toward Developing Perfect Serodiagnostic Target Using Trichomonas vaginalis as a Model
Time : 09:40-10:25
Dr.Alderete received his PhD at the age of 28 years from Kansas University in 1978 did postdoctoral research at University of North Carolina-Chapel Hill. He was at the University of Texas Health Science Center at San Antonio for 30 years before taking a positon at Washington State University. He has published 140 scientific articles and 63 book chapters, invited articles, and press releases. His work has been presented at 158 scientific conferences. He has served in National Institutes of Health Study Sections, Boards of Scientific Counselors, National Advisory Councils, and several National Academy of Medicine panels.
Trichomonas vaginalis causes the number one, non-viral sexually transmitted infection world-wide. A rapid, sensitive and accurate Point-of-Care serodiagnostic is needed for screening both women and men, and such a diagnostic will permit determination of the true incidence and prevalence of this STI responsible for significant adverse health outcomes. The availability of sera from women and men patients and uninfected controls allowed us to identify epitopes to immunogenic proteins of T. vaginalis. I reasoned that knowledge of epitope amino acid sequences could permit the construction of novel, chimeric recombinant proteins that would be a perfect target for a serum IgG diagnostic for both women and men. The metabolic enzymes fructose-1,6-bisphosphate aldolase (A), α-enolase (E), and glyceraldehyde-3-phosphate dehydro-genase (G) are immunogenic, and serum IgG antibody to these proteins is detected after this STI. Some epitopes of these enzymes have little or no sequence identity to other eukaryotes, yeasts, and microbial pathogens. We constructed a chimeric recombinant String-Of-Epitopes (SOE) protein consisting of 15-mer peptides within which are epitopes of A, E, and G unique to this STI agent. This chimeric protein referred to as AEG::SOE2 was detected by ELISA with highly reactive sera of women and men, but not control, negative serum lacking antibody to trichomonad proteins. This approach lends itself to the creation of highly specific immunogenic targets for detection of serum IgG antibody in patients. Given that the epitopes are highly immunogenic and elicit antibody, such targets may also be future subunit vaccine candidates.
Keynote: Enzyme immobilization using glutaraldehyde: taking advantages of the versatility of the method
Time : 10:50-11:35
Roberto Fernandez-Lafuente has completed his PhD at UAM and postdoctoral studies from UCL-London. He is the leading the “Optimization of biocatalysts and bioprocess” group at ICP-CSIC, a premier Bio-Soft service organization. He has published more than 390 papers in ISI Journal, with an H index of 68 (Scopus) and more than 2200 citations/year, he is coauthor of 20 patents and co-supervisor of 18 doctoral thesis. He is associated editor of Process Biochemistry and has been serving as an editorial board member of more than 20 journals (e.g., Molecules, Enzyme and Microbial Technology, Journal of Biotechnology, Molecular Catalysis, etc.).
Glutaraldehyde is among the most employed reagents in the preparation of immobilized enzyme biocatalysts. Usually, a support containing primary amino groups is used. This way, this support is actually a heterofunctional one with ion exchange capacity and hydrophobic groups, and also bearing chemical reactivity that can generate covalent bonds. For this reason, glutaraldehyde may immobilize enzyme by very different reasons, biocatalysts with very different activity/stability properties. For example: Adsorption of the enzymes on the aminated support via ion exchange followed by treatment with glutaraldehyde. Use of preactivated supports at low ionic strength, where the first step of the immobilization remains the ion exchange. Use of preactivated supports at high ionic stregnth to avoid ion exchange as first step of the immobilization, forcing the covalent attachment as the first immobilization step, that may depend on the immobilization pH. Lipases will be treated as a particlar case, as they can become interfacially adosrbed on the activated supports.