Method for detecting cell glycosylation level

A glycosylation, horizontal technology, applied in the field of cell glycosylation detection and analysis, can solve the problem of limited conjugation of cells and unevenness

Inactive Publication Date: 2015-04-22
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, inherent problems exist, including the need to stain the cells resulting in limited conjugation of the cells to the lectin binding site.

Method used

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  • Method for detecting cell glycosylation level
  • Method for detecting cell glycosylation level
  • Method for detecting cell glycosylation level

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Embodiment 1

[0023] A method for detecting cell glycosylation level in this embodiment comprises the following steps:

[0024] 1) Drop 100 μL of chloroauric acid solution with a concentration of 0.25 mol / L into 10 mL of CTAB solution with a concentration of 0.1 mol / L, and then add 600 μL of 0.01 mol / L sodium borohydride solution, mix for two minutes to make seed solution by reducing chloroauric acid with sodium borohydride, and maintain the seed solution at 25°C;

[0025] 2) Add 700μL 4mmol / L silver nitrate, 4mL 0.24mol / L chloroauric acid and 275μL 80mmol / L vitamin C to the CTAB solution with a volume of 10mL and a concentration of 0.1mol / L to make a growth solution;

[0026] 3) Add about 60 μL of the seed solution obtained in step 1) to the growth solution obtained in step 2), stir slowly, and react for several hours to obtain gold nanorods. Centrifuge the obtained gold nanorods at 8000rmp for 30 minutes After that, remove the CTAB in the upper layer, resuspend the obtained gold nanorods...

Embodiment 2

[0034] A method for detecting cell glycosylation level in this embodiment comprises the following steps:

[0035] 1) Drop 100 μL of chloroauric acid solution with a concentration of 0.20 mol / L into 10 mL of CTAB solution with a concentration of 0.1 mol / L, and then add 600 μL of 0.01 mol / L sodium borohydride solution, mix for two minutes to make seed solution by reducing chloroauric acid with sodium borohydride, and maintain the seed solution at 25°C;

[0036] 2) Add 600μL 4mmol / L silver nitrate, 4mL 0.25mol / L chloroauric acid and 280μL 70mmol / L vitamin C to the CTAB solution with a volume of 10mL and a concentration of 0.1mol / L to make a growth solution;

[0037] 3) Add about 60 μL of the seed solution obtained in step 1) to the growth solution obtained in step 2), stir slowly, and react for several hours to obtain gold nanorods. Centrifuge the obtained gold nanorods at 7500rmp for 30 minutes After that, remove the CTAB in the upper layer, resuspend the obtained gold nanorods...

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Abstract

The invention provides a method for detecting cell glycosylation level by utilizing gold nano rods. The method comprises the following steps: dripping chloroauric acid into a CTAB solution, and then adding a sodium borohydride solution to prepare a seed solution; adding silver nitrate, chloroauric acid and vitamin C into the CTAB solution to prepare a growth solution; dripping the seed solution into the growth solution, and centrifuging after reaction to obtain gold nano rods; adding COOH-PEG-SH and sulfydryl malic acid into the gold nano rods for reaction to realize bio-functionalization activation of the gold nano rods so as to prepare an activated gold nano rod solution; connecting WGA agglutinin to the activated gold nano rods to obtain the activated gold nano rods provided with the WDA agglutinin and used for detecting the glycosylation level; and detecting corresponding glycosylated proteins in a cell lysis buffer by observing changes of an ultraviolet-visible absorption spectrum.

Description

technical field [0001] The invention relates to the field of cell glycosylation detection and analysis, in particular to the detection of the glycosylation level of activated gold nanorods. Background technique [0002] Glycosyls play an important role in physiological and biological events related to intracellular and extracellular signaling, and also have an impact on the interaction of cancer metastasis, and may also play an important role in the pathogenesis of many diseases. Previous studies have shown abnormal changes in glycosylation levels, such as highly expressed glycans in some cancer cells, indicating that the detection of glycosylation levels is promising for biomarkers in early diagnosis, prognosis, and detection in cancer targeted therapy and treatment basis and contributions. [0003] Although fluorescence-based biolabeling and imaging applications are commonly used to study glycobiological events, the complex glycan structures consist of rapid and sensitive...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552G01N21/31G01N21/33
Inventor 汪敏王禹陈刘英帅
Owner SOUTHWEST UNIV
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