Inhibitors in Commercially Available 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonate) Affect Enzymatic Assays

Zhang, Yifei; Hess, Henry

doi:10.1021/acs.analchem.9b04751

Cited by 12 publications

(7 citation statements)

References 71 publications

(109 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Above results indicated that ABTS was not a good color reagent for monitoring peroxidation in basic conditions and over long periods of time in the presence of excess H 2 O 2 . Other studies also reported that impurities of commercial ABTS could inhibit enzymes in the reaction [25] . Another color substrate, tetramethylbenzidine (TMB), was also assessed and was found to be unstable under basic pH conditions and H 2 O 2 .…”

Section: Resultsmentioning

confidence: 99%

“…Other studies also reported that impurities of commercial ABTS could inhibit enzymes in the reaction. [25] Another color substrate, tetramethylbenzidine (TMB), was also assessed and was found to be unstable under basic pH conditions and H 2 O 2 . The color signal from TMB was quickly saturated or even depleted after a few minutes of GQH-catalyzed reaction due to the substrate instability and catalyst inactivation (Figure S5).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Inactivation Kinetics of G‐Quadruplex/Hemin Complex and Optimization for More Reliable Catalysis

Carlo

2022

ChemPlusChem

View full text Add to dashboard Cite

Reliable catalysis is critical for the synthesis of various chemicals, molecular sensing and biomedicine. G-quadruplex/Hemin (GQH) complex, a peroxidase-mimicking DNAzyme, has been widely used in various publications. However, a concern exists about the unstable kinetics of GQH-catalyzed peroxidation. This work investigates several factors that result in the inactivation of GQH and the signal degradation during long reaction periods, including pH, buffer component, the selection of substrate and the oxidation damage of cofactor. Using colori-metric and fluorescent assays, GQH was found to be highly unstable under basic conditions with 50 % of GQH activity lost within 2 minutes at high H 2 O 2 concentrations. Appropriate conditions and substrates are suggested for accurately characterizing GQH-catalyzed reactions, as well as optimization to improve the catalytic reliability, such as the use of polyhistidine and cascade reactions. These results could be useful for GQHrelated applications.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Inactivation Kinetics of G‐Quadruplex/Hemin Complex and Optimization for More Reliable Catalysis

Carlo

2022

ChemPlusChem

View full text Add to dashboard Cite

show abstract

“…[ 21 ] Alternatively, it could also oxidize the colorless ABTS to the green ABTS + under the existence of H 2 O 2 (Figure S17b, Supporting Information). [ 22 ] As shown in Figure 4d, PABP/HRP would not catalyze TMB or ABTS to blue (TMBH) or green (ABTS + ) products. While incubation with GSH could recover the enzymes’ activities, which catalyzed substrates to products with corresponding colors.…”

Section: Resultsmentioning

confidence: 99%

Crosslinked Protein Delivery Strategy with Precise Activity Regulation Properties for Cancer Therapy and Gene Editing

Liu

Zhu

Sun

et al. 2022

Adv Healthcare Materials

View full text Add to dashboard Cite

Protein drugs hold tremendous promise for therapeutic applications due to their direct and superior pharmacological effects. However, protein drugs can be degraded in blood stream and unable to cross many physical barriers to exert therapeutic effect. Degradable synthetic crosslinking is a versatile strategy to enhance the stability of the nanoparticle in a complex physiological medium and is helpful to get through physical barriers. Herein, crosslinked polypeptide (PABP) composed of poly-amino acids including cystine, tyrosine, lysine, ketal bridge, and polyethylene glycol (PEG) is modularly explored and synthesized for protein delivery. Notably, plasma membrane V-ATPase is the particular pathway which induces the macropinocytosis of the inner peptide analogous core (PAB/protein) after the outer PEG shell disassociation at tumor intercellular sites. In addition, PABP/protein achieves proteins' activity shielding in systemic circulation and recovery in tumor cytoplasm precisely. In application, PABP/RNase-A shows satisfying tumor accumulation and antineoplastic efficacy. More importantly, PABP/Cas9 + small guide RNA displays obvious gene editing efficiency. The crosslinked protein delivery strategy not only makes the accurate protein transport and activity regulation possible but also is promising in paving the way for clinical translation of protein drugs.

show abstract

“…With the exceptions mentioned below, all chemicals used were the same as those reported in Ghećzy et al 42 In the case of hydrogen peroxide (H 2 O 2 ) and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS 2− (NH 4 + ) 2 ), new batches of the same product were used; no significant difference was observed with respect to the ones used before. 47 2.1.1. Enzymes and Enzyme Substrates.…”

Section: Methodsmentioning

confidence: 99%

Controllable Enzyme Immobilization via Simple and Quantitative Adsorption of Dendronized Polymer–Enzyme Conjugates Inside a Silica Monolith for Enzymatic Flow-Through Reactor Applications

Ghéczy

Szymańska

et al. 2022

ACS Omega

View full text Add to dashboard Cite

Although many different methods are known for the immobilization of enzymes on solid supports for use in flow-through applications as enzyme reactors, the reproducible immobilization of predetermined amounts of catalytically active enzyme molecules remains challenging. This challenge was tackled using a macro- and mesoporous silica monolith as a support and dendronized polymer–enzyme conjugates. The conjugates were first prepared in an aqueous solution by covalently linking enzyme molecules and either horseradish peroxidase (HRP) or bovine carbonic anhydrase (BCA) along the chains of a water-soluble second-generation dendronized polymer using an established procedure. The obtained conjugates are stable biohybrid structures in which the linking unit between the dendronized polymer and each enzyme molecule is a bisaryl hydrazone (BAH) bond. Quantitative and reproducible enzyme immobilization inside the monolith is possible by simply adding a defined volume of a conjugate solution of a defined enzyme concentration to a dry monolith piece of the desired size. In that way, (i) the entire volume of the conjugate solution is taken up by the monolith piece due to capillary forces and (ii) all conjugates of the added conjugate solution remain stably adsorbed (immobilized) noncovalently without detectable leakage from the monolith piece. The observed flow-through activity of the resulting enzyme reactors was directly proportional to the amount of conjugate used for the reactor preparation. With conjugate solutions consisting of defined amounts of both types of conjugates, the controlled coimmobilization of the two enzymes, namely, BCA and HRP, was shown to be possible in a simple way. Different stability tests of the enzyme reactors were carried out. Finally, the enzyme reactors were applied to the catalysis of a two-enzyme cascade reaction in two types of enzymatic flow-through reactor systems with either coimmobilized or sequentially immobilized BCA and HRP. Depending on the composition of the substrate solution that was pumped through the two types of enzyme reactor systems, the coimmobilized enzymes performed significantly better than the sequentially immobilized ones. This difference, however, is not due to a molecular proximity effect with regard to the enzymes but rather originates from the kinetic features of the cascade reaction used. Overall, the method developed for the controllable and reproducible immobilization of enzymes in the macro- and mesoporous silica monolith offers many possibilities for systematic investigations of immobilized enzymes in enzymatic flow-through reactors, potentially for any type of enzyme.

show abstract

Inhibitors in Commercially Available 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonate) Affect Enzymatic Assays

Cited by 12 publications

References 71 publications

Inactivation Kinetics of G‐Quadruplex/Hemin Complex and Optimization for More Reliable Catalysis

Inactivation Kinetics of G‐Quadruplex/Hemin Complex and Optimization for More Reliable Catalysis

Crosslinked Protein Delivery Strategy with Precise Activity Regulation Properties for Cancer Therapy and Gene Editing

Controllable Enzyme Immobilization via Simple and Quantitative Adsorption of Dendronized Polymer–Enzyme Conjugates Inside a Silica Monolith for Enzymatic Flow-Through Reactor Applications

Contact Info

Product

Resources

About