Protein tyrosine phosphatase 1B;
Lateral flow immunoassay;
Peptide functionalized gold nanoparticles;
Living cell;
PTP1B;
ASSAY;
KINASE;
CANCER;
D O I:
10.1016/j.ab.2022.114671
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
In this work, a lateral flow immunoassay (LFIA) with peptide functionalized gold nanoparticles (termed as biotin-ppeptide-AuNPs) has been developed for rapid, semi-quantitative detection of PTP1B activity without using any sophisticated equipment. In this method, the anti-phosphotyrosine (anti-pY) monoclonal antibody and streptavidin were used as test line and control line, respectively. The biotin-ppeptide-AuNPs contain 10% biotinylated peptide ligand carry a motif SDGHEpYIYVDP with pY (phosphotyrosine) and 90% pentapeptide (CALNN) ligand, which are used as PTP1B substrates and LFIA labelling probes. The experimental results demonstrate that the as-proposed LFIA with biotin-ppeptide-AuNPs exhibits a wide linear range (from 50 ng/mL to 10 mu g/mL), a relatively low limit of detection (LOD, 44 ng/mL), and good specificity. In addition, the LFIA with biotin-(p)peptide-AuNPs has been successfully used to evaluate activity levels of PTP1B in four cell lysates and the detection results exhibit a consistent trend with that of commercial kit.
机构:
Univ Nebraska, Dept Chem, Lincoln, NE 68588 USA
Southeast Missouri State Univ, Dept Chem & Phys, Cape Girardeau, MO 63701 USAUniv Nebraska, Dept Chem, Lincoln, NE 68588 USA
Casey, Garrett R.
Stains, Cliff, I
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机构:
Univ Nebraska, Dept Chem, Lincoln, NE 68588 USA
Univ Virginia, Dept Chem, Charlottesville, VA 22904 USA
Univ Nebraska, Nebraska Ctr Integrated Biomol Commun, Lincoln, NE 68588 USA
Univ Nebraska Med Ctr, Canc Genes & Mol Regulat Program, Fred & Pamela Buffet & Canc Ctr, Omaha, NE 68198 USA
Univ Virginia, Canc Ctr, Charlottesville, VA 22904 USAUniv Nebraska, Dept Chem, Lincoln, NE 68588 USA
机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Chen, Queting
Li, Yong
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机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Li, Yong
Li, Zhong
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h-index: 0
机构:
Hebei Univ, Affiliated Hosp, Dept Surg Oncol, Baoding 071000, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Li, Zhong
Zhao, Qun
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h-index: 0
机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Zhao, Qun
Fan, Liqiao
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h-index: 0
机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
机构:
Univ Nebraska, Dept Chem, Lincoln, NE 68588 USA
Southeast Missouri State Univ, Dept Chem & Phys, Cape Girardeau, MO 63701 USAUniv Nebraska, Dept Chem, Lincoln, NE 68588 USA
Casey, Garrett R.
Stains, Cliff, I
论文数: 0引用数: 0
h-index: 0
机构:
Univ Nebraska, Dept Chem, Lincoln, NE 68588 USA
Univ Virginia, Dept Chem, Charlottesville, VA 22904 USA
Univ Nebraska, Nebraska Ctr Integrated Biomol Commun, Lincoln, NE 68588 USA
Univ Nebraska Med Ctr, Canc Genes & Mol Regulat Program, Fred & Pamela Buffet & Canc Ctr, Omaha, NE 68198 USA
Univ Virginia, Canc Ctr, Charlottesville, VA 22904 USAUniv Nebraska, Dept Chem, Lincoln, NE 68588 USA
机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Chen, Queting
Li, Yong
论文数: 0引用数: 0
h-index: 0
机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Li, Yong
Li, Zhong
论文数: 0引用数: 0
h-index: 0
机构:
Hebei Univ, Affiliated Hosp, Dept Surg Oncol, Baoding 071000, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Li, Zhong
Zhao, Qun
论文数: 0引用数: 0
h-index: 0
机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China
Zhao, Qun
Fan, Liqiao
论文数: 0引用数: 0
h-index: 0
机构:
Hebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R ChinaHebei Med Univ, Affiliated Hosp 4, Dept Gen Surg, Shijiazhuang 050011, Hebei, Peoples R China