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Original Research

A highly sensitive and selective viral protein detection method based on RNA oligonucleotide nanoparticle

, , &
Pages 323-329 | Published online: 22 Apr 2010

Figures & data

Figure 1 Representative scheme for the detection of viral protein on chip. A) Scheme for viral protein assay on chip. B) Procedure for viral protein assay.

Figure 1 Representative scheme for the detection of viral protein on chip. A) Scheme for viral protein assay on chip. B) Procedure for viral protein assay.

Figure 2 The conjugation pattern on 2% agarose gel electrophoresis was examined at UV excitation wavelength (345 nm), free QDs (lane 1), QDs-NS5Bnor RNA oligonucleotide (lane 2), and QDs-NS5Bmut RNA oligonucleotide (lane 3). Carboxyl-terminated QDs are conjugated with RNA oligonucleotide by an amine-carboxyl reaction.

Abbreviations: NS5B, nonstructural protein 5B; QDs, quantum dots; UV, ultraviolet.

Figure 2 The conjugation pattern on 2% agarose gel electrophoresis was examined at UV excitation wavelength (345 nm), free QDs (lane 1), QDs-NS5Bnor RNA oligonucleotide (lane 2), and QDs-NS5Bmut RNA oligonucleotide (lane 3). Carboxyl-terminated QDs are conjugated with RNA oligonucleotide by an amine-carboxyl reaction.Abbreviations: NS5B, nonstructural protein 5B; QDs, quantum dots; UV, ultraviolet.

Figure 3 Characterization of QDs by TEM analysis. A) Free QDs. B) QDs-conjugated RNA oligonucleotide.

Abbreviations: QDs, quantum dots; TEM, transmission electron microscopy.

Figure 3 Characterization of QDs by TEM analysis. A) Free QDs. B) QDs-conjugated RNA oligonucleotide.Abbreviations: QDs, quantum dots; TEM, transmission electron microscopy.

Figure 4 Purification of HCV-NS5B viral protein. The SDS-PAGE 10% gel showing NS5B protein with his-tag. M, protein marker; lane 1, before induction form of NS5B; lane 2, total form of NS5B; lane 3, soluble form of NS5B; lane 4, his-Tag form of NS5B.

Abbreviations: HCV, hepatitis C virus; NS5B, non-structural protein 5B; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.

Figure 4 Purification of HCV-NS5B viral protein. The SDS-PAGE 10% gel showing NS5B protein with his-tag. M, protein marker; lane 1, before induction form of NS5B; lane 2, total form of NS5B; lane 3, soluble form of NS5B; lane 4, his-Tag form of NS5B.Abbreviations: HCV, hepatitis C virus; NS5B, non-structural protein 5B; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.

Figure 5 A, B) Selective specificity of HCV-NS5B viral protein by QDs-conjugated RNA oligonucleotide. The NS5Bnor and NS5Bmut RNA oligonucleotides were bound on an immobilized viral protein chip, respectively. C, D) Effect of HCV-NS5B viral protein concentrations on binding assay.

Abbreviations: HCV, hepatitis C virus; NS5B, non-structural protein 5B; NS5Bnor, normal NS5B; NS5Bmut, mutant NS5B; QDS, quantum dots.

Figure 5 A, B) Selective specificity of HCV-NS5B viral protein by QDs-conjugated RNA oligonucleotide. The NS5Bnor and NS5Bmut RNA oligonucleotides were bound on an immobilized viral protein chip, respectively. C, D) Effect of HCV-NS5B viral protein concentrations on binding assay.Abbreviations: HCV, hepatitis C virus; NS5B, non-structural protein 5B; NS5Bnor, normal NS5B; NS5Bmut, mutant NS5B; QDS, quantum dots.