oriented immobilization
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2022 ◽  
Author(s):  
Bruna Coelho de Andrade ◽  
Adriano Gennari ◽  
Gaby Renard ◽  
Edilson Valmir Benvenutti ◽  
Jocelei Maria Chies ◽  
...  

The Analyst ◽  
2022 ◽  
Author(s):  
Masumi Iijima ◽  
Yuki Yamada ◽  
Hideo Nakano ◽  
Tsutomu Nakayama ◽  
Shun'ichi Kuroda

The oriented immobilization of sensing molecules (e.g., IgGs, receptors, lectins, and DNA aptamers) on sensor chips is particularly important for maximizing the sensitivity and target-binding capacity of biosensors.


Biosensors ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 466
Author(s):  
Ieva Šakinytė ◽  
Marius Butkevičius ◽  
Vidutė Gurevičienė ◽  
Jonita Stankevičiūtė ◽  
Rolandas Meškys ◽  
...  

As electrode nanomaterials, thermally reduced graphene oxide (TRGO) and modified gold nanoparticles (AuNPs) were used to design bioelectrocatalytic systems for reliable D-tagatose monitoring in a long-acting bioreactor where the valuable sweetener D-tagatose was enzymatically produced from a dairy by-product D-galactose. For this goal D-fructose dehydrogenase (FDH) from Gluconobacter industrius immobilized on these electrode nanomaterials by forming three amperometric biosensors: AuNPs coated with 4-mercaptobenzoic acid (AuNP/4-MBA/FDH) or AuNPs coated with 4-aminothiophenol (AuNP/PATP/FDH) monolayer, and a layer of TRGO on graphite (TRGO/FDH) were created. The immobilized FDH due to changes in conformation and spatial orientation onto proposed electrode surfaces catalyzes a direct D-tagatose oxidation reaction. The highest sensitivity for D-tagatose of 0.03 ± 0.002 μA mM–1cm–2 was achieved using TRGO/FDH. The TRGO/FDH was applied in a prototype bioreactor for the quantitative evaluation of bioconversion of D-galactose into D-tagatose by L-arabinose isomerase. The correlation coefficient between two independent analyses of the bioconversion mixture: spectrophotometric and by the biosensor was 0.9974. The investigation of selectivity showed that the biosensor was not active towards D-galactose as a substrate. Operational stability of the biosensor indicated that detection of D-tagatose could be performed during six hours without loss of sensitivity.


2021 ◽  
pp. 106609
Author(s):  
Mariia Vorobii ◽  
Rita Teixeira-Santos ◽  
Luciana C. Gomes ◽  
Manuela Garay-Sarmiento ◽  
Anna M. Wagner ◽  
...  

2021 ◽  
Vol 216 ◽  
pp. 108788
Author(s):  
Wenfeng Zhou ◽  
Yuan Rao ◽  
Wei Zhuang ◽  
Lei Ge ◽  
Rijia Lin ◽  
...  

2021 ◽  
Author(s):  
Zhiquan Wang ◽  
Deyi Wu ◽  
Xinze Wang ◽  
Hainan Kong

Abstract Surfactants, especially non-ionic surfactants, play an important role in the preparation of nanocarriers and can also promote the enzymatic hydrolysis of lignocellulose. A broad overview of the current status of surfactants on the immobilization of cellulase is provided in this review. In addition, the restricting factors in cellulase immobilization in the complex multiphase hydrolysis system are discussed, including the carrier structure characteristics, solid-solid contact obstacles, external diffusion resistance, limited recycling frequency, and invalid combination of enzyme active centers. Furthermore, promising prospects of cellulase-oriented immobilization are proposed, including the hydrophilic-hydrophobic interaction of surfactants and cellulase in the oil-water reaction system, the reversed micelle system of surfactants, and the possible oriented immobilization mechanism.


2021 ◽  
Vol 21 (5) ◽  
pp. 2834-2840
Author(s):  
José M. Abad ◽  
Sara Puertas ◽  
Daniel Pérez ◽  
Christian Sánchez-Espinel

Antibody-functionalized gold nanoparticle constitutes a powerful interface biosystem for biomedical applications where the properties of gold nanoparticles and the specificity of antibody–antigen interactions are combined. This study provides insight into the key factors for the development of antibody functionalized gold nanoparticles focusing on the immobilization of the antibody. Here, we address an oriented antibody immobilization procedure on gold nanoparticles. It comprises chelatemodified gold nanoparticles that are designed for oriented immobilization of IgG antibodies (end on spatial orientation) through the metal-chelation to histidine-rich metal binding site in the heavy chain (Fc) of the antibody.


Sensors ◽  
2021 ◽  
Vol 21 (8) ◽  
pp. 2702
Author(s):  
Madalina M. Barsan ◽  
Caroline G. Sanz ◽  
Melania Onea ◽  
Victor C. Diculescu

A dual strategy for the electrochemical detection for 20S proteasome (20S) is proposed, based on the oriented immobilization of a capture monoclonal antibody (Abβ) on a self-assembled monolayer of 4-mercaptophenylboronic acid (4-MPBA) on gold electrodes, which led to the Au/4-MPBA/Abβ immunosensor. The methodology comprises the correlation of 20S concentration with (i) its proteolytic activity toward the Z-LLE-AMC substrate, using the Au/4-MPBA/Abβ/20S, and (ii) the enzymatic activity of an alkaline phosphatase (AlkP) from the AlkP-labeled secondary antibody (Abcore-AlkP), which involves the conversion of aminophenylphosphate to the electroactive aminophenol using Au/4-MPBA/Abβ/20S/Abcore-AlkP. The step-by-step construction of the immunosensor and the interactions at its surface were evaluated by surface plasmon resonance and gravimetric analysis with quartz crystal microbalance, showing a high affinity between both antibodies and 20S. Morphological analysis by scanning electron microscopy demonstrated a pattern of parallel lines upon immobilization of Abβ on 4-MPBA and morphological changes to a well-organized granular structure upon binding of 20S. A voltametric and impedimetric characterization was performed after each step in the immunosensor construction. The two detection strategies were evaluated. It was shown that the immunosensor responds linearly with 20S concentration in the range between 5 and 100 µg mL−1, which corresponds to proteasome levels in serum in the case of diverse pathological situations, and LoD values of 1.4 and 0.2 µg mL−1 were calculated for the detection strategies. The immunosensor was applied to the detection of 20S in serum samples with recovery values ranging from 101 to 103%.


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