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Amplified voltammetric detection of glycoproteins using 4-mercaptophenylboronic acid/biotin-modified multifunctional gold nanoparticles as labels.

Liu L, Xing Y, Zhang H, Liu R, Liu H, Xia N - Int J Nanomedicine (2014)

Bottom Line: The MBA-biotin-AuNPs facilitated the attachment of streptavidin-conjugated alkaline phosphatase for the production of electroactive p-aminophenol from p-aminophenyl phosphate substrate.A detection limit of 8 fmol L(-1) for rHuEPO detection was achieved.Other glycosylated and non-glycosylated proteins, such as horseradish peroxidase, prostate specific antigen, metallothionein, streptavidin, and thrombin showed no interference in the detection assay.

View Article: PubMed Central - PubMed

Affiliation: College of Chemistry and Chemical Engineering, Anyang Normal University, Anyang, Henan, People's Republic of China ; College of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan, People's Republic of China.

ABSTRACT
Ultrasensitive detection of protein biomarkers is essential for early diagnosis and therapy of many diseases. Glycoproteins, differing from other types of proteins, contain carbohydrate moieties in the oligosaccharide chains. Boronic acid can form boronate ester covalent bonds with diol-containing species. Herein, we present a sensitive and cost-effective electrochemical method for glycoprotein detection using 4-mercaptophenylboronic acid (MBA)/biotin-modified gold nanoparticles (AuNPs) (MBA-biotin-AuNPs) as labels. To demonstrate the feasibility and sensitivity of this method, recombinant human erythropoietin (rHuEPO) was tested as a model analyte. Specifically, rHuEPO was captured by the anti-rHuEPO aptamer-covered electrode and then derivatized with MBA-biotin-AuNPs through the boronic acid-carbohydrate interaction. The MBA-biotin-AuNPs facilitated the attachment of streptavidin-conjugated alkaline phosphatase for the production of electroactive p-aminophenol from p-aminophenyl phosphate substrate. A detection limit of 8 fmol L(-1) for rHuEPO detection was achieved. Other glycosylated and non-glycosylated proteins, such as horseradish peroxidase, prostate specific antigen, metallothionein, streptavidin, and thrombin showed no interference in the detection assay.

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Ultraviolet-visible absorption spectra of the citrate-stabilized AuNPs (curve a) and the synthesized MBA-biotin-AuNPs (curve b). The inset shows the transmission electron microscopy image of the MBA-biotin-AuNPs.Abbreviations: Abs, absorbance; AuNP, gold nanoparticle; MBA-biotin-AuNP, 4-mercaptophenylboronic acid/biotin-modified gold nanoparticle.
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f2-ijn-9-2619: Ultraviolet-visible absorption spectra of the citrate-stabilized AuNPs (curve a) and the synthesized MBA-biotin-AuNPs (curve b). The inset shows the transmission electron microscopy image of the MBA-biotin-AuNPs.Abbreviations: Abs, absorbance; AuNP, gold nanoparticle; MBA-biotin-AuNP, 4-mercaptophenylboronic acid/biotin-modified gold nanoparticle.

Mentions: Modification of AuNPs with multiple functionalities can provide more flexibility for multiplexing in bioanalytical application. For example, Kong et al35 reported the colorimetric dopamine detection using MBA-DSP-AuNPs (AuNPs modified with MBA and dithiobis[succinimidylpropionate]). Such bifunctional AuNPs can doubly recognize dopamine by reacting with the diol and amine groups. AuNPs modified with CALNN peptide are water-soluble and extremely stable.36 Wang et al37 demonstrated that multifunctional AuNPs capped with CALNNGK(biotin)G peptide and DNA targets are readily obtained in one step through the ligand-exchange reaction. Such functionality facilitates SA to be absorbed on the surface of AuNPs through the strong biotin–SA interaction. In the present work, the bifunctional AuNPs capped with MBA and CALNNGK(biotin)G groups were prepared by the formation of the Au–S covalent bond. The synthesized MBA-biotin-AuNPs were characterized by UV-visible spectroscopy. As shown in Figure 2, the MBA-biotin-AuNPs displayed a characteristic UV-visible absorption spectrum with a plasmon band at 520 nm (curve b). Such absorption was ascribed to the surface plasmon resonance of the AuNPs (curve a). This demonstrated that the synthesized MBA-biotin-AuNPs were stable and monodisperse. The result was also confirmed by transmission electron microscopy, as shown in the inset of Figure 2.


Amplified voltammetric detection of glycoproteins using 4-mercaptophenylboronic acid/biotin-modified multifunctional gold nanoparticles as labels.

Liu L, Xing Y, Zhang H, Liu R, Liu H, Xia N - Int J Nanomedicine (2014)

Ultraviolet-visible absorption spectra of the citrate-stabilized AuNPs (curve a) and the synthesized MBA-biotin-AuNPs (curve b). The inset shows the transmission electron microscopy image of the MBA-biotin-AuNPs.Abbreviations: Abs, absorbance; AuNP, gold nanoparticle; MBA-biotin-AuNP, 4-mercaptophenylboronic acid/biotin-modified gold nanoparticle.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4043723&req=5

f2-ijn-9-2619: Ultraviolet-visible absorption spectra of the citrate-stabilized AuNPs (curve a) and the synthesized MBA-biotin-AuNPs (curve b). The inset shows the transmission electron microscopy image of the MBA-biotin-AuNPs.Abbreviations: Abs, absorbance; AuNP, gold nanoparticle; MBA-biotin-AuNP, 4-mercaptophenylboronic acid/biotin-modified gold nanoparticle.
Mentions: Modification of AuNPs with multiple functionalities can provide more flexibility for multiplexing in bioanalytical application. For example, Kong et al35 reported the colorimetric dopamine detection using MBA-DSP-AuNPs (AuNPs modified with MBA and dithiobis[succinimidylpropionate]). Such bifunctional AuNPs can doubly recognize dopamine by reacting with the diol and amine groups. AuNPs modified with CALNN peptide are water-soluble and extremely stable.36 Wang et al37 demonstrated that multifunctional AuNPs capped with CALNNGK(biotin)G peptide and DNA targets are readily obtained in one step through the ligand-exchange reaction. Such functionality facilitates SA to be absorbed on the surface of AuNPs through the strong biotin–SA interaction. In the present work, the bifunctional AuNPs capped with MBA and CALNNGK(biotin)G groups were prepared by the formation of the Au–S covalent bond. The synthesized MBA-biotin-AuNPs were characterized by UV-visible spectroscopy. As shown in Figure 2, the MBA-biotin-AuNPs displayed a characteristic UV-visible absorption spectrum with a plasmon band at 520 nm (curve b). Such absorption was ascribed to the surface plasmon resonance of the AuNPs (curve a). This demonstrated that the synthesized MBA-biotin-AuNPs were stable and monodisperse. The result was also confirmed by transmission electron microscopy, as shown in the inset of Figure 2.

Bottom Line: The MBA-biotin-AuNPs facilitated the attachment of streptavidin-conjugated alkaline phosphatase for the production of electroactive p-aminophenol from p-aminophenyl phosphate substrate.A detection limit of 8 fmol L(-1) for rHuEPO detection was achieved.Other glycosylated and non-glycosylated proteins, such as horseradish peroxidase, prostate specific antigen, metallothionein, streptavidin, and thrombin showed no interference in the detection assay.

View Article: PubMed Central - PubMed

Affiliation: College of Chemistry and Chemical Engineering, Anyang Normal University, Anyang, Henan, People's Republic of China ; College of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan, People's Republic of China.

ABSTRACT
Ultrasensitive detection of protein biomarkers is essential for early diagnosis and therapy of many diseases. Glycoproteins, differing from other types of proteins, contain carbohydrate moieties in the oligosaccharide chains. Boronic acid can form boronate ester covalent bonds with diol-containing species. Herein, we present a sensitive and cost-effective electrochemical method for glycoprotein detection using 4-mercaptophenylboronic acid (MBA)/biotin-modified gold nanoparticles (AuNPs) (MBA-biotin-AuNPs) as labels. To demonstrate the feasibility and sensitivity of this method, recombinant human erythropoietin (rHuEPO) was tested as a model analyte. Specifically, rHuEPO was captured by the anti-rHuEPO aptamer-covered electrode and then derivatized with MBA-biotin-AuNPs through the boronic acid-carbohydrate interaction. The MBA-biotin-AuNPs facilitated the attachment of streptavidin-conjugated alkaline phosphatase for the production of electroactive p-aminophenol from p-aminophenyl phosphate substrate. A detection limit of 8 fmol L(-1) for rHuEPO detection was achieved. Other glycosylated and non-glycosylated proteins, such as horseradish peroxidase, prostate specific antigen, metallothionein, streptavidin, and thrombin showed no interference in the detection assay.

Show MeSH
Related in: MedlinePlus