Enzymatic Deposition of Silver Particles for Detecting Protease Activity

Ding, Xiaokang; Yang, Kun‐Lin

doi:10.1002/ppsc.201400107

Cited by 7 publications

(6 citation statements)

References 38 publications

(17 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ascorbic acid (AA) is a commonly used reducing agent for the growth of metal (eg, Au and Ag) nanoparticles. 34,35 Free Cu 2+ ion can initiate the catalyzed oxidation of AA in the presence of O 2 . 36 In this process, AA is oxidized into dehydroascorbate by free Cu 2+ ion; then, the resultant Cu + ion is subsequently oxidized into Cu 2+ by O 2 .…”

Section: Introductionmentioning

confidence: 99%

Gold nanoparticle-based colorimetric method for the detection of prostate-specific antigen

Xia¹,

Deng²,

Wang³

et al. 2018

IJN

View full text Add to dashboard Cite

BackgroundProstate-specific antigen (PSA), a serine protease, is a biomarker for preoperative diagnosis and screening of prostate cancer and monitoring of its posttreatment.MethodsIn this work, we reported a colorimetric method for clinical detection of PSA using gold nanoparticles (AuNPs) as the reporters. The method is based on ascorbic acid (AA)-induced in situ formation of AuNPs and Cu2+-catalyzed oxidation of AA. Specifically, HAuCl4 can be reduced into AuNPs by AA; Cu2+ ion can catalyze the oxidation of AA by O2 to inhibit the formation of AuNPs. In the presence of the PSA-specific peptide (DAHSSKLQLAPP)-modified gold-coated magnetic microbeads (MMBs; denoted as DAHSSKLQLAPP-MMBs), complexation of Cu2+ by the MMBs through the DAH–Cu2+ interaction depressed the catalyzed oxidation of AA and thus allowed for the formation of red AuNPs. However, once the peptide immobilized on the MMB surface was cleaved by PSA, the DAHSSKLQ segment would be released. The resultant LAPP fragment remaining on the MMB surface could not sequestrate Cu2+ to depress its catalytic activity toward AA oxidation. Consequently, no or less AuNPs were generated.ResultsThe linear range for PSA detection was found to be 0~0.8 ng/mL with a detection limit of 0.02 ng/mL. Because of the separation of cleavage step and measurement step, the interference of matrix components in biological samples was avoided.ConclusionThe high extinction coefficient of AuNPs facilitates the colorimetric analysis of PSA in serum samples. This work is helpful for designing of other protease biosensors by matching specific peptide substrates.

show abstract

Section: Introductionmentioning

confidence: 99%

Gold nanoparticle-based colorimetric method for the detection of prostate-specific antigen

Xia¹,

Deng²,

Wang³

et al. 2018

IJN

View full text Add to dashboard Cite

show abstract

“…However, with the growing interest for understanding protease activities as potential drug targets or biomarkers for diseases, there has been a growing emphasis on the need for more extensive studies to be carried out in a complex matrix or real biological samples. Such a matrix can cause unforeseen interferences to Trypsin (3 nM) Cao et al 66 Electrochemical R||GGLAC Trypsin (0.6 nM) Park et al 149 Electrochemical GPR||(AP) b Trypsin (∼0.02 pM) Ding and Yang 157 Colorimetric PPLR||INR||HILTR||GGG-(biotin) Trypsin (0.4 nM) Bi et al 100 LC CDR||VYIHPFHLK Trypsin (2 nM) Chen and Yang 101 LC CK||GSNR||TR||IDEGNQR||ATR||MLGGKETSAAKLKR||K||YWW Trypsin (0.04 nM) CL||SEL||DDRADAL||QAGASQF||ESSAAKL||KRKY||W||W||KNL||K Chymotrypsin (4 pM)…”

Section: Discussionmentioning

confidence: 99%

“…For example, Ding et al immobilised biotin-labelled peptides (PPLR||INR||HILTR|| GGG-(biotin)) on a glass surface, and then conjugated the peptides to Sav-ALP. 157 The enzyme (ALP) was able to remove a phosphate group from ascorbic acid 2-phosphate and produced ascorbic acid, which reduced silver ions to silver particles. Based on this principle, they reported an LOD of 0.4 nM for trypsin.…”

Section: Enzyme-linked Peptide Protease Assaysmentioning

confidence: 99%

See 1 more Smart Citation

Recent developments in protease activity assays and sensors

Ong

Yang

2017

Analyst

Self Cite

104

View full text Add to dashboard Cite

Proteases play a pivotal role in regulating important physiological processes from food digestion to blood clotting. They are also important biomarkers for many diseases such as cancers. The importance of proteases has led to extensive efforts in the screening of proteases and their inhibitors as potential drug molecules. For example, human immunodeficiency virus (HIV) patients have been treated with HIV-1 protease inhibitors to prolong the life expectancy of patients. Such a close relationship between diseases and proteases provides a strong motivation for developing sensitive, selective, and robust protease assays and sensors, which can be exploited to discover new proteases and inhibitors. In this aspect, protease assays based on levels of proteolytic activities are more relevant than protease affinity assays such as immunoassays. In this review, recent developments of protease activity assays based on different detection principles are discussed and compared. For homogenous assays, fluorescence-based techniques are the most popular due to their high sensitivity and quantitative results. However, homogeneous assays have limited multiplex sensing capabilities. In contrast, heterogeneous assays can be employed to detect multiple proteases simultaneously, given the microarray technology that is already available. Among them, electrochemical methods, surface spectroscopy techniques, and enzyme-linked peptide protease assays are commonly used. Finally, recent developments in liquid crystal (LC)-based protease assays and their applications for detecting proteases and their inhibitors are discussed.

show abstract

“…Proteases are able to catalyze the hydrolysis of the amide bonds in proteins or peptides and thereafter trigger series of cascading biochemistry reactions to regulate numerous biological and physiological processes. − The dysfunction or overactivity of proteases is associated with many pathological processes, such as arthritis, cirrhosis, Alzheimer’s disease, cardiovascular disease, and tumor invasion and metastasis . Therefore, a variety of techniques were developed for the detection and quantification of the protease activities. − Although the high-performance-liquid-chromatography or mass-spectroscopy − -based techniques are able to detect the peptide fragments cleaved by the proteases, they are limited by their bulky and complicated instrumentation. Alternatively, several state-of-the-art protease assays were reported based on conjugated polymers, − aggregation of gold nanoparticles, − quantum dots, − Förster resonance energy transfer, , and aggregation-induced emission effect .…”

Section: Introductionmentioning

confidence: 99%

Rational Design of Peptide-Functionalized Poly(Methacrylic Acid) Brushes for On-Chip Detection of Protease Biomarkers

Nizam

Ding

2017

ACS Biomater. Sci. Eng.

Self Cite

View full text Add to dashboard Cite

There is an increasing demand for developing new materials and approaches for rapid and sensitive detection of protease biomarkers. Herein, the poly(methacrylic acid) (PMAA) brushes were synthesized from silica nanoparticles via surface-initiated atom transfer radical polymerization (ATRP), and flexibly functionalized with different fluorescein-labeled peptides, serving as the substrates for protease assay. To facilitate the point-of-care detection of protease, polyacrylamide gel pad arrays were fabricated to allow permeation of fluorescein-labeled peptide fragments cleaved from the PMAA brushes. This experimental setup enables an on-chip protease assay with an adequate limit of detection (LOD) for detecting trypsin in a buffer solution (3.9 pM) or in serum (1.4 nM) and good specificity for differentiation of trypsin and chymotrypsin. By using this experimental setup, matrix metalloproteinase-2 and matrix metalloproteinase-9 can be detected with LODs of 2.5 nM and 3.3 nM, respectively. Moreover, by introducing an adamantine (Ad) motif to the side-chain of the peptide fragment and β-cyclodextrin (β-CD) groups to the gel pad matrix, a 2.2-fold lower LOD was achieved for the detection of trypsin (1.8 pM) due to the supramolecular self-assembly of Ad and β-CD. Given the advances in the ease of sample handling, this rational design of peptide-functionalized PMAA brushes could be useful for on-chip detection of protease biomarkers or the screening of potential protease inhibitors.

show abstract

Enzymatic Deposition of Silver Particles for Detecting Protease Activity

Cited by 7 publications

References 38 publications

Gold nanoparticle-based colorimetric method for the detection of prostate-specific antigen

Gold nanoparticle-based colorimetric method for the detection of prostate-specific antigen

Recent developments in protease activity assays and sensors

Rational Design of Peptide-Functionalized Poly(Methacrylic Acid) Brushes for On-Chip Detection of Protease Biomarkers

Contact Info

Product

Resources

About