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1.
Front Immunol ; 14: 1276950, 2023.
Article in English | MEDLINE | ID: mdl-38179057

ABSTRACT

Introduction: This study evaluated the immune response to a multiepitope recombinant chimeric protein (CHIVAX) containing B- and T-cell epitopes of the SARS-CoV-2 spike's receptor binding domain (RBD) in a translational porcine model for pre-clinical studies. Methods: We generated a multiepitope recombinant protein engineered to include six coding conserved epitopes from the RBD domain of the SARS-CoV-2 S protein. Pigs were divided into groups and immunized with different doses of the protein, with serum samples collected over time to determine antibody responses by indirect ELISA and antibody titration. Peptide recognition was also analyzed by Western blotting. A surrogate neutralization assay with recombinant ACE2 and RBDs was performed. Intranasal doses of the immunogen were also prepared and tested on Vietnamese minipigs. Results: When the immunogen was administered subcutaneously, it induced specific IgG antibodies in pigs, and higher doses correlated with higher antibody levels. Antibodies from immunized pigs recognized individual peptides in the multiepitope vaccine and inhibited RBD-ACE2 binding for five variants of concern (VOC). Comparative antigen delivery methods showed that both, subcutaneous and combined subcutaneous/intranasal approaches, induced specific IgG and IgA antibodies, with the subcutaneous approach having superior neutralizing activity. CHIVAX elicited systemic immunity, evidenced by specific IgG antibodies in the serum, and local mucosal immunity, indicated by IgA antibodies in saliva, nasal, and bronchoalveolar lavage secretions. Importantly, these antibodies demonstrated neutralizing activity against SARS-CoV-2 in vitro. Discussion: The elicited antibodies recognized individual epitopes on the chimeric protein and demonstrated the capacity to block RBD-ACE2 binding of the ancestral SARS-CoV-2 strain and four VOCs. The findings provide proof of concept for using multiepitope recombinant antigens and a combined immunization protocol to induce a neutralizing immune response against SARS-CoV-2 in the pig translational model for preclinical studies.


Subject(s)
COVID-19 , Vaccines , Swine , Animals , Humans , Immunity, Mucosal , COVID-19/prevention & control , COVID-19 Vaccines , SARS-CoV-2 , Angiotensin-Converting Enzyme 2 , Swine, Miniature , Epitopes, T-Lymphocyte , Immunoglobulin A , Immunoglobulin G
2.
Int J Biol Macromol ; 183: 1723-1731, 2021 Jul 31.
Article in English | MEDLINE | ID: mdl-34051250

ABSTRACT

This work focused on studying the mineral composition, morphology, thermal, structural, and pasting properties of isolated plantain starch. Plantain starch is rich in K, and other ions as Mg, Ca, P, and Si were found. This starch exhibits lenticular, elliptical, and semispherical morphologies. Two endothermal events present in the thermogram were identified as the hexagonal and orthorhombic solvation. C-type starch formed by hexagonal and orthorhombic nanocrystal was completely indexed. The ash content showed the presence of calcium phosphate (KCaP2O7), Calcium Magnesium Phosphate (Ca2.71Mg0.29(PO4)2), and silicon oxide (SiO2). The pasting profile of this starch behaves between a custard and a hydrogel. Scanning electron microscopy of the lyophilized samples along pasting profile confirms that the shear and van der Walls forces and slurry morphology govern the pasting profile changes.


Subject(s)
Flour/analysis , Musa/chemistry , Starch/chemistry , Calorimetry, Differential Scanning , Microscopy, Electron, Scanning , Molecular Structure , Shear Strength , X-Ray Diffraction
3.
Food Chem ; 220: 490-497, 2017 Apr 01.
Article in English | MEDLINE | ID: mdl-27855929

ABSTRACT

This research studied the influence of the germ components on the physicochemical properties of cooked corn and nixtamalized corn flours as a function of the calcium hydroxide content (from 0 to 2.1 w/w) and steeping time (between 0 and 9h). A linear relationship was found between calcium content in germ and steeping time used during nixtamalization process. X-ray diffraction analysis showed that calcium carbonate is formed into the germ structure to 2.1 w/w of calcium hydroxide and 9h steeping time. The presence of the germ improves the development of peak viscosity in flours, and it is related to the increases in calcium concentration in germ and the formation of amylose-lipid complexes. No significant changes were observed in palmitic, stearic, oleic and linoleic acids of corn oil. The levels of further corn oil deterioration were 2.1 w/w of calcium hydroxide concentration and 9h of steeping time.


Subject(s)
Corn Oil/analysis , Flour/analysis , Food Handling/methods , Viscosity , Zea mays/chemistry , Amylose/chemistry , Amylose/metabolism , Calcium/metabolism , Cooking/methods , Fatty Acids/analysis , Germination , X-Ray Diffraction
4.
Int J Biol Macromol ; 91: 106-14, 2016 Oct.
Article in English | MEDLINE | ID: mdl-27211296

ABSTRACT

This work is focused on the understanding of the apparent viscosity profile of corn starch, in terms of the physicochemical and morphological changes that take place during the thermal profile of starch-water suspension to its respective gel formation. A mathematical model was used to obtain the experimental operating conditions that satisfy the Froude number. Freeze drying samples are studied in different stages along the pasting profile. Changes in the structural properties of the samples are studied using X-ray diffraction, and the morphological changes are followed using scanning electron microscopy, differential scanning calorimetry was used to analyze the thermal changes in starch. The changes in the pasting profile are associated with structural, thermal,and morphological changes of the system and the analysis of the physicochemical transformation that occur during the pasting profile are explained. The finding in this work does not show evidence of gel retrogradation at the end of the cooling process.


Subject(s)
Starch/chemistry , Temperature , Amylose/analysis , Ointments , Viscosity , Water/chemistry
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