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1.
Biosensors (Basel) ; 13(7)2023 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-37504072

RESUMO

Diagnostic techniques based on biomolecules have application potential that can be realized in many fields, such as disease diagnosis, bioprocess imaging, food/beverage industries, and environmental pollutant imaging. Successful surface immobilization of biomolecules is critical to increasing the stabilization, sensitivity, and selectivity of biomolecules used in bioassay systems. Nanofibers are good candidates for the immobilization of biomolecules owing to many advantages such as morphology and pore size. In this study, montmorillonite (MMT) clay is modified with poly(amidoamine) (PAMAM) generation 3 (PAMAMG3) and added to polystyrene (PS) solutions, following which PS/MMT-PAMAMG3 nanofibers are obtained using the electrospinning method. The nanofibers are obtained by testing PS% (wt%) and MMT-PAMAMG3% (wt%) ratios and characterized with scanning electron microscopy. Antiserum amyloid A antibody (Anti-SAA) is then conjugated to the nanofibers on the electrode surface via covalent bonds using a zero-length cross linker. Finally, the obtained selective surface is used for electrochemical determination of serum amyloid A (SAA) levels. The linear range of PS/MMT-PAMAM/Anti-SAA is between 1 and 200 ng/mL SAA, and the detection limit is 0.57 ng/mL SAA. The applicability of PS/MMT-PAMAMG3/Anti-SAA is investigated by taking measurements in synthetic saliva and serum both containing SAA.


Assuntos
Técnicas Biossensoriais , Nanofibras , Poliestirenos , Argila , Proteína Amiloide A Sérica , Nanofibras/química , Imunoensaio
2.
Biosensors (Basel) ; 13(4)2023 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-37185504

RESUMO

Herein, dendrimer-modified montmorillonite (Mt)-decorated poly-Ɛ-caprolactone (PCL) and chitosan (CHIT)-based nanofibers were prepared. Mt was modified with a poly(amidoamine) generation 1 (PAMAMG1) dendrimer, and the obtained PAMAMG1-Mt was incorporated into the PCL-CHIT nanofiber's structure. The PCL-CHIT/PAMAMG1-Mt nanofibers were conjugated with glutamate oxidase (GluOx) to design a bio-based detection system for monosodium glutamate (MSG). PAMAMG1-Mt was added to the PCL-CHIT backbone to provide a multipoint binding side to immobilize GluOx via covalent bonds. After the characterization of PCL-CHIT/PAMAMG1-Mt/GluOx, it was calibrated for MSG. The linear ranges were determined from 0.025 to 0.25 mM MSG using PCL-CHIT/Mt/GluOx and from 0.0025 to 0.175 mM MSG using PCL-CHIT/PAMAMG1-Mt/GluOx (with a detection limit of 7.019 µM for PCL-CHIT/Mt/GluOx and 1.045 µM for PCL-CHIT/PAMAMG1-Mt/GluOx). Finally, PCL-CHIT/PAMAMG1-Mt/GluOx was applied to analyze MSG content in tomato soup without interfering with the sample matrix, giving a recovery percentage of 103.125%. Hence, the nanofiber modification with dendrimer-intercalated Mt and GluOx conjugation onto the formed nanocomposite structures was performed, and the PCL-CHIT/PAMAMG1-Mt/GluOx system was successfully developed for MSG detection.


Assuntos
Técnicas Biossensoriais , Quitosana , Dendrímeros , Nanofibras , Glutamato de Sódio , Nanofibras/química , Eletrodos , Quitosana/química
3.
Biosens Bioelectron ; 109: 286-294, 2018 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-29573728

RESUMO

The construction and biofunctionalization of the poly (Ɛ-caprolactone) (PCL)-chitosan (CHIT) nanofibrous mats, which included Polyamidoamine (PAMAM) dendrimer modified montmorillonite (Mt), for the cell adhesion and electrochemical cytosensing were accomplished in this report. After the intercalation of the PAMAM generation zero dendrimer into the Mt, PAMAM-Mt decorated PCL-CHIT electrospun nanofibers were formed. The addition of PAMAM caused the decrease of contact angle of PCL-CHIT nanofibers. The covalent immobilization of a tripeptide namely Arginylglycylaspartate (RGD) on both the PCL-CHIT/Mt and PCL-CHIT/PAMAM-Mt surface was carried out. U87-MG and HaCaT (negative control) cell lines were incubated on the PCL-CHIT/Mt/RGD and PCL-CHIT/PAMAM-Mt/RGD. The proliferation studies and imaging of the cells were carried out on these fibers. Finally, electrochemical measurements were performed after each modification step by differential pulse/cyclic voltammetry and electrochemical impedance spectroscopy. U87-MG cells were grown better than HaCaT cells on the PCL-CHIT/PAMAM-Mt/RGD surfaces. To the best of our knowledge, there is no study that developed electrochemical cytosensor using electrospun nanofibers as a cell adhesion platform.


Assuntos
Técnicas Biossensoriais , Proliferação de Células , Nanofibras/química , Engenharia Tecidual/métodos , Bentonita/química , Adesão Celular , Quitosana/química , Dendrímeros/química , Poliésteres/química , Alicerces Teciduais/química
4.
J Chromatogr A ; 1361: 43-52, 2014 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-25130089

RESUMO

Clay material plays an important role in the transport and retention of many compounds in the soil, therefore, clay based sorbents are promising alternatives for selective sorption of organic pollutants. In the present work, different chain length ionic liquids (ILs) namely, 1-methyl-3-octyl-imidazolium bromide, 1-methyl-3-undecyl-imidazolium bromide and 1-methyl-3-octadecyl-imidazolium bromide were intercalated in the galleries of montmorillonite (MMT) clay. Then, this novel nanofiller surface was utilized in micro extraction of estrogenic hormones for the first time. A fast procedure where sonication-assisted emulsification microextraction combined with vortex assisted micro-solid phase extraction (µ-SPE) was developed for the LC-MS/MS analysis of estrone (E1), 17ß-estradiol (E2), estriol (E3) and ethynylestradiol (EE2). The parameters related to the µ-SPE procedure namely; pH, sorbent amount, extraction solvent type and volume, sonication and vortex time, sample volume and salt effect on the extraction efficiency were screened by applying Plackett-Burmann design. The selected parameters were then optimized by using Box-Behnken design. The method was validated for the determination of estrogenic hormone residues in river water samples. Linear calibration plots were obtained for all hormones whose regression coefficients were larger than 0.98. RSD values were found less than 10% for three levels of concentration. LOD levels were calculated as; 0.012, 0.062, 0.018 and 0.693 ng L(-1) for E1, E2, E3 and EE2, respectively. Recovery values were calculated in the range of 86.9-97.7%. Considering large sample volumes required for attaining low limits of these hormones, present method provides an ease for analyst as 10 mL of the sample is adequate for achieving the same sensitivity.


Assuntos
Silicatos de Alumínio/química , Cromatografia Líquida de Alta Pressão/métodos , Líquidos Iônicos/química , Rios/química , Microextração em Fase Sólida/métodos , Esteroides/isolamento & purificação , Espectrometria de Massas em Tandem/métodos , Argila , Íons , Limite de Detecção , Sonicação
5.
Macromol Biosci ; 11(1): 141-50, 2011 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-20945305

RESUMO

Hydrophobic- and/or hydrophilic-polymer-grafted PDVB microspheres are synthesized by the combination of hydrobromination and click-chemistry processes. The modified-PDVB microspheres and the intermediates at various stages of synthesis are characterized using GPC, ¹H NMR and FTIR spectroscopy and TGA analysis. Use of the microspheres as a support matrix for reversible protein immobilization via adsorption is investigated. The system parameters such as the adsorption conditions (i.e., enzyme concentration, medium pH) and desorption are studied and evaluated with regards to the biocatalytic activity and adsorption capacity.


Assuntos
Lacase/química , Microesferas , Estirenos/química , Adsorção , Agaricus/química , Química Click , Ensaios Enzimáticos , Enzimas Imobilizadas , Halogenação , Ácido Bromídrico/química , Concentração de Íons de Hidrogênio
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