Electrochemical detection of protein kinase activity based on carboxypeptidase Y digestion triggered signal amplification

Yin, Huanshun; Wang, Xinxu; Guo, Yunlong; Zhou, Yunlei; Ai, Shiyun

doi:10.1016/j.bios.2014.11.014

Cited by 30 publications

(12 citation statements)

References 31 publications

(28 reference statements)

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“…The process for culturing of human hepatic cell line L-02 and human hepatoma carcinoma cell line HepG2 and extracting total protein from these cells were according to our previous report …”

Section: Methodsmentioning

confidence: 99%

Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO₂/g-C₃N₄, PAMAM Dendrimer, and Alkaline Phosphatase

et al. 2017

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A novel photoelectrochemical (PEC) assay is developed for sensitive detection of protein kinase A (PKA) activity based on PKA-catalyzed phosphorylation reaction in solution and signal amplification strategy triggered by PAMAM dendrimer and alkaline phosphatase (ALP). In this strategy, it is noteworthy at this point that PKA phosphorylation was achieved in solution instead of on the surface of the electrode, which has advantages of the good contact in reactants and simple experimental procedure. For immobilizing the phosphorylated peptide (P-peptide) on electrode surface, graphite-like carbon nitride (g-CN) and titanium dioxide (TiO) complex is synthesized and characterized, which plays a significant role for TiO conjugating phosphate groups and g-CN providing PEC signal. Subsequently, PAMAM dendrimer and ALP can be captured on P-peptide and TiO/g-CN modified ITO electrode via interaction between the -COOH groups on the surface of PAMAM dendrimer and the -NH groups of peptide and ALP, which can lead to the increase of ALP amount on the modified electrode surface assisted with the PAMAM dendrimer. As a result, the amount of ALP catalyzes of L-ascorbic acid 2-phosphate trisodium salt (AAP) to produce electron donor of ascorbic acid (AA), resulting in an increased photocurrent. The proposed detection assay displays high selectivity and low detection limit of 0.048 U/mL (S/N = 3) for PKA activity. This biosensor can also be applied for the evaluation of PKA inhibition and PKA activity assay in cell samples. Therefore, the fabricated PEC biosensor is potentionally well in PKA activity detection and inhibitor screening.

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“…The process for culturing of human hepatic cell line L-02 and human hepatoma carcinoma cell line HepG2 and extracting total protein from these cells were according to our previous report …”

Section: Methodsmentioning

confidence: 99%

Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO₂/g-C₃N₄, PAMAM Dendrimer, and Alkaline Phosphatase

et al. 2017

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“…The addition of subsequent layers on the sensor surface acts as a barrier for the interfacial electron transfer reaction of the [Fe(CN) 6 ] 3−/4− process, which results in a decrease in DPV current response. [18][19][20][21][22] To demonstrate the applicability of the method, we selected ovalbumin 9 as a model protein. This protein has multiple lectin binding sites and, since it is not present in humans, it is suitable as an external protein for quantification in the background of human serum samples.…”

Section: Resultsmentioning

confidence: 99%

“…MCH is a short chain alkane with a thiol and a hydroxyl end that allows it to adsorb in the gold surface at an upright conformation with the thiol end at the bottom and the hydroxyl group at the top. This orientation of MCH perfectly blocks larger nonspecific molecules such as DNA or proteins 20,25,26 to bind either with the exposed gold surface or to the MCH molecule itself but still allows the Fe(CN) 6 ] 3−/4− solution to come through the small gap to reach the gold surface for generating Faradaic current. The MCH concentration was optimized through comparison of current reduction values upon addition of either lectin or streptavidin control (Fig.…”

Section: Resultsmentioning

confidence: 99%

Electrochemical detection of protein glycosylation using lectin and protein–gold affinity interactions

Yadav

Carrascosa

Sina

et al. 2016

Analyst

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We report a new method for the electrochemical detection of glycosylation on proteins, which relies on lectin-protein interaction on a bare gold electrode. The target protein isolated by immunoaffinity is directly adsorbed onto a gold surface and its glycosylation status is retrieved by subsequent addition of specific lectins. The adsorption and subsequent recognition process is monitored electrochemically in the presence of [Fe(CN)6](3-/4-) redox system. By decoupling target protein capture from glycosylation read-out steps, this approach circumvents unwanted antibody-lectin crosstalk while enabling specific glycosylation detection of a glycoprotein in serum-spiked samples in less than 1 h.

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“…Gold nanoparticles electrodeposited onto gold electrodes have also been used as supports for biosensor development. An electrochemical sensor for protein casein kinase II (CK2) activity was constructed using gold nanoparticles electrodeposited onto a gold electrode [18]. The electrodeposition was carried out at −0.2 V (versus Ag/AgCl) for 250 s in a solution of three mM of HAuCl 4 containing 0.1 M of KNO 3 .…”

Section: Deposition At a Constant Potential For A Fixed Time Periodmentioning

confidence: 99%

Biosensor Applications of Electrodeposited Nanostructures

Stine

2019

Applied Sciences

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The development of biosensors for a range of analytes from small molecules to proteins to oligonucleotides is an intensely active field. Detection methods based on electrochemistry or on localized surface plasmon responses have advanced through using nanostructured electrodes prepared by electrodeposition, which is capable of preparing a wide range of different structures. Supported nanoparticles can be prepared by electrodeposition through applying fixed potentials, cycling potentials, and fixed current methods. Nanoparticle sizes, shapes, and surface densities can be controlled, and regular structures can be prepared by electrodeposition through templates. The incorporation of multiple nanomaterials into composite films can take advantage of the superior and potentially synergistic properties of each component. Nanostructured electrodes can provide supports for enzymes, antibodies, or oligonucleotides for creating sensors against many targets in areas such as genomic analysis, the detection of protein antigens, or the detection of small molecule metabolites. Detection can also be performed using electrochemical methods, and the nanostructured electrodes can greatly enhance electrochemical responses by carefully designed schemes. Biosensors based on electrodeposited nanostructures can contribute to the advancement of many goals in bioanalytical and clinical chemistry.

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Electrochemical detection of protein kinase activity based on carboxypeptidase Y digestion triggered signal amplification

Cited by 30 publications

References 31 publications

Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO₂/g-C₃N₄, PAMAM Dendrimer, and Alkaline Phosphatase

Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO₂/g-C₃N₄, PAMAM Dendrimer, and Alkaline Phosphatase

Electrochemical detection of protein glycosylation using lectin and protein–gold affinity interactions

Biosensor Applications of Electrodeposited Nanostructures

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Electrochemical detection of protein kinase activity based on carboxypeptidase Y digestion triggered signal amplification

Cited by 30 publications

References 31 publications

Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO2/g-C3N4, PAMAM Dendrimer, and Alkaline Phosphatase

Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO2/g-C3N4, PAMAM Dendrimer, and Alkaline Phosphatase

Electrochemical detection of protein glycosylation using lectin and protein–gold affinity interactions

Biosensor Applications of Electrodeposited Nanostructures

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Product

Resources

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Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO₂/g-C₃N₄, PAMAM Dendrimer, and Alkaline Phosphatase

Enhanced Photoelectrochemical Method for Sensitive Detection of Protein Kinase A Activity Using TiO₂/g-C₃N₄, PAMAM Dendrimer, and Alkaline Phosphatase