Evaluation of click chemistry microarrays for immunosensing of alpha-fetoprotein (AFP).

The level of cancer biomarkers in cells, tissues or body fluids can be used for the prediction of the presence of cancer or can even indicate the stage of the disease. Alpha-fetoprotein (AFP) is the most commonly used biomarker for early screening and diagnosis of hepatocellular carcinoma (HCC). Here, a combination of three techniques (click chemistry, the biotin-streptavidin-biotin sandwich strategy and the use of antigen-antibody interactions) were combined to implement a sensitive fluorescent immunosensor for AFP detection. Three types of functionalized glasses (dibenzocyclooctyne- (DBCO-), thiol- and epoxy-terminated surfaces) were biotinylated by employing the respective adequate click chemistry counterparts (biotin-thiol or biotin-azide for the first class, biotin-maleimide or biotin-DBCO for the second class and biotin-amine or biotin-thiol for the third class). The anti-AFP antibody was immobilized on the surfaces via a biotin-streptavidin-biotin sandwich technique. To evaluate the sensing performance of the differently prepared surfaces, fluorescently labeled AFP was spotted onto them via microchannel cantilever spotting (µCS). Based on the fluorescence measurements, the optimal microarray design was found and its sensitivity was determined.

Site-Specific Surface Functionalization via Microchannel Cantilever Spotting (µCS): Comparison between Azide-Alkyne and Thiol-Alkyne Click Chemistry Reactions.

Different types of click chemistry reactions are proposed and used for the functionalization of surfaces and materials, and covalent attachment of organic molecules. In the present work, two different catalyst-free click approaches, namely azide-alkyne and thiol-alkyne click chemistry are studied and compared for the immobilization of microarrays of azide or thiol inks on functionalized glass surfaces. For this purpose, the surface of glass is first functionalized with dibenzocyclooctyne-acid (DBCO-acid), a cyclooctyne with a carboxyl group. Then, the DBCO-terminated surfaces are functionalized via microchannel cantilever spotting with different fluorescent and nonfluorescent azide and thiol inks. Although both routes work reliably for surface functionalization, the protein binding experiments reveal that using a thiol-alkyne route will obtain the highest surface density of molecular immobilization in such spotting approaches. The obtained achievements and results from this work can be used for design and manufacturing of microscale patterns suitable for biomedical and biological applications.

Investigation of gelatin/multi-walled carbon nanotube nanocomposite films as packaging materials.

Gelatin composite films were prepared from gelatin solutions (10% w/v) containing multi-walled carbon nanotubes (MWCNT, 0.5, 1, 1.5, and 2% w/w gelatin) as nanofiller. The water solubility, water swelling, water uptake, water vapor permeability (WVP), mechanical, and antibacterial properties of the films were examined. Water solubility, water swelling, water uptake, and WVP for gelatin films were 45 ± 1%, 821 ± 42%, 45 ± 1.1%, and 0.4 ± 0.022 g mm/m(2) kPa h, respectively. Incorporation of MWCNT caused a significant decrease in water solubility, water swelling, water uptake, and WVP. Gelatin/MWCNT films containing 1-1.5% MWCNT showed the lowest water vapor transmission. Tensile strength, elongation at break, and Young's modulus for gelatin films were 13.4 ± 1.2 MPa, 95 ± 5%, and 45.4 ± 7 MPa, respectively. Incorporation of MWCNT caused a significant increase in tensile strength and decrease in the elongation at break. The largest mechanical strength was found at 1.5% MWCNT. All gelatin/MWCNT films showed significant antibacterial activities against both gram-positive and gram-negative bacteria. Our results suggest that the gelatin/MWCNT composites films could be used as a very attractive alternative to traditional materials for different biomedical and food applications.

Mechanical, physical, antioxidant, and antimicrobial properties of gelatin films incorporated with thymol for potential use as nano wound dressing.

UNLABELLED: The aim of this study was to determine the properties of gelatin films incorporated with thymol. Gelatin films were prepared from gelatin solutions (10% w/v) containing thymol (1, 2, 4, and 8% w/w), glycerol (25% w/w) as plasticizer, and glutaraldehyde (2% w/w) as cross-linker. Cross-likened films showed higher tensile strength, higher elongation at break, lower Young's modulus, lower water solubility, lower swelling, lower water uptake, and lower water vapor permeability. Incorporation of thymol caused a significant decrease in tensile strength, increase in elongation at break, decrease in Young's modulus, increase in water solubility, decrease in swelling and water uptake, and increase in water vapor permeability slightly. The films incorporated with thymol exhibited excellent antioxidant and antibacterial properties. The antibacterial activity of the films containing thymol was greatest against Staphylucoccus aureus followed by Bacillus subtilis followed by Escherichia coli and then by Pseudomonas aeruginosa. Thus, gelatin films-containing thymol can be used as safe and effective source of natural antioxidant and antimicrobial agents with the purpose of evaluating their potential use as modern nano wound dressing. PRACTICAL APPLICATION: This study clearly demonstrates the potential of gelatin films incorporated with thymol as natural antioxidant and antimicrobial nano film. Such antimicrobial films exhibited excellent mechanical, physical, and water activities and could be used as antibacterial nano wound dressing against wounds burn pathogens.