Direct and Selective Immobilization of Proteins by Means of an Inorganic Material-Binding Peptide: Discussion on Functionalization in the Elongation to Material-Binding Peptide
ABSTRACT Using an artificial peptide library, we have identified a peptide with affinity for ZnO materials that could be used to selectively accumulate ZnO particles on polypropylene-gold plates. In this study, we fused recombinant green fluorescent protein (GFP) with this ZnO-binding peptide (ZnOBP) and then selectively immobilized the fused protein on ZnO particles. We determined an appropriate condition for selective immobilization of recombinant GFP, and the ZnO-binding function of ZnOBP-fused GFP was examined by elongating the ZnOBP tag from a single amino acid to the intact sequence. The fusion of ZnOBP with GFP enabled specific adsorption of GFP on ZnO substrates in an appropriate solution, and thermodynamic studies showed a predominantly enthalpy-dependent electrostatic interaction between ZnOBP and the ZnO surface. The ZnOBP's binding affinity for the ZnO surface increased first in terms of material selectivity and then in terms of high affinity as the GFP-fused peptide was elongated from a single amino acid to intact ZnOBP. We concluded that the enthalpy-dependent interaction between ZnOBP and ZnO was influenced by the presence of not only charged amino acids but also their surrounding residues in the ZnOBP sequence.
- SourceAvailable from: Fernando Ponz
[Show abstract] [Hide abstract]
- "This has induced many ongoing efforts aimed to tackle this problem using very different strategies (Moyle et al., 2006; Saenz et al., 2009; Seo et al., 2009; Villa et al., 2011). For antibody or protein binding on solid phases, peptides have the clear disadvantage over larger proteins of their low affinity for the most common solid phases used for macromolecule immobilization, a problem which has also led to specific and often sophisticated approaches to get around it (Li et al., 2011; Palmieri et al., 1995; Yokoo et al., 2010). So, efforts for more intense developments of peptide-based technologies in immunology need to deal with these important hurdles. "
ABSTRACT: Elongated and flexuous recombinant nanoparticles were derived from Turnip mosaic virus to be used as bioscaffolds for increased peptide immunogenicity and peptide-specific antibody sensing. For this purpose, a 20-amino acid peptide derived from human vascular endotelial growth factor receptor 3 (VEGFR-3) was fused to the N-terminal region of Turnip mosaic virus coat protein (CP) by genetic insertion. The insertion was between codons corresponding to the first and second amino acids of the CP in two versions of a previously reported virus-derived vector. Systemic infections of two genetic constructs were achieved in two different plant hosts. The construct proved stable upon successive passages and generated virus nanoparticles identifiable under the electron microscope. The chimeric structures held the VEGFR-3 peptide. Purified VER3 nanoparticles were used to immunize mice, whose sera showed log increases of antibodies against the VEGFR-3 peptide when compared with mice immunized with peptide alone, thus providing the first quantitative data on the potential of elongated flexuous viruses for peptide immunogenicity increases. Purified VER3 nanoparticles also showed log increases in their ability to detect VER3 antibodies in sera, when used as reagents in ELISA assays, an application also used here for the first time.Journal of Biotechnology 09/2013; 168(4). DOI:10.1016/j.jbiotec.2013.09.002 · 2.88 Impact Factor
Conference Paper: Filter spectral selection for 3 wavelength optical pyrometry[Show abstract] [Hide abstract]
ABSTRACT: The filter spectral selection problem for a 3-wavelength optical pyrometer is analyzed in connection with the nonideal spectral distribution of infrared radiation from a high temperature body, taking into account spectral emissivity from a specific material. The level of spectral radiance is evaluated in order to maximize the ratio between the signals from three wavelengths, two at the time. A mathcad optimization procedure for filter spectral selection is proposedSemiconductor Conference, 2001. CAS 2001 Proceedings. International; 02/2001
Conference Paper: Design of wavelength dependent optical attenuators[Show abstract] [Hide abstract]
ABSTRACT: In this paper simulated annealing algorithm (SA) and coupled mode theory (CMT) are combined to design wavelength dependant optical attenuator by using long period gratings (LPGs). Both principles and method are presented. Various wavelength dependant attenuators are designed. The results show a good agreement with the required specification.Lasers and Electro-Optics, 2001. CLEO/Pacific Rim 2001. The 4th Pacific Rim Conference on; 02/2001