Abstract
Polyvinyl alcohol (PVA), PVA crosslinked with glutaraldehyde hydrogels (PVA/GA), PVA with tetraethylorthosilicate (PVA/TEOS) and PVA/GA/TEOS hybrids with recombinant MPB70 protein (rMPB70) incorporated were chemically characterized by Fourier transform infrared spectroscopy (FTIR). FTIR spectra of PVA hydrogel samples showed the absorption regions of the specific chemical groups associated with poly(vinyl alcohol) (-OH, -CO, -CH2) and PVA/GA confirming the formation of crosslinked hydrogel (duplet -CH). It was observed C-H broad alkyl stretching band (n = 2850-3000 cm-1) and typical strong hydroxyl bands for free alcohol (nonbonded -OH stretching band at n = 3600-3650 cm-1), and hydrogen bonded band (n = 3200-3570 cm-1). The most important vibration bands related to silane alcoxides have been verified on FTIR spectra of PVA/TEOS and PVA/GA/TEOS hybrids (Si-O-Si, n = 1080 and n = 450 cm-1; Si-OH, n = 950 cm-1). FTIR spectra of f PVA hydrogel with rMPB70 incorporated have indicated the specific groups usually found in protein structures, such as amides I, II and III, at 1680-1620 cm-1, 1580-1480 cm-1 and 1246 cm-1, respectively. These results have given strong evidence that recombinant protein rMPB70 was successfully adsorbed in the hydrogels and hybrids networks. These PVA based hydrogels and hybrids were further used in immunological assays (Enzyme-Linked Immunosorbent Assay - ELISA). Tests were performed to detect antibodies against rMPB70 protein in serum samples from bovines that were positive in the tuberculin test. Corresponding tests were carried out without PVA samples in microtiter plates as control. Similar results were found for commercially available microplates and PVA based hydrogels and hybrids developed in the present work regarding to immunoassay sensitivity and specificity response.
Highlights
The biomaterials and Bioengineering research field have broadened in the last 3 decades, including replacement of diseased or damaged parts, assist in healing, correct and improve functional abnormality, drug delivery systems, immunological kits and biosensors[1,2,3,4,5,6,7]
Fourier transform infrared spectroscopy (FTIR) spectrum in Figure 3b is associated with Polyvinyl alcohol (PVA) crosslinked by glutaraldehyde (PVA/GA)
It can be observed that two important peaks at ν = 2860 and 2730 cm-1 of C–H stretching are related to aldehydes, a duplet absorption with peaks attributed to the alkyl chain[7,16,17]
Summary
The biomaterials and Bioengineering research field have broadened in the last 3 decades, including replacement of diseased or damaged parts, assist in healing, correct and improve functional abnormality, drug delivery systems, immunological kits and biosensors[1,2,3,4,5,6,7]. The basic properties of PVA are dependent on the degree of polymerization or on the degree of hydrolysis. It has been widely used in adhesives, emulsificants, in the textile and paper industry applications and in the attainment of amphiphilic membranes for enzyme immobilization[6]. Hybrids would combine properties of organic polymers with ceramics These different components can be mixed at length scales ranging from nanometer to micrometer, in virtually any ratio leading to the so-called hybrid organic-inorganic materials.
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