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A kind of cyclometal platinum complex and its preparation method and its application as protein staining agent

A technology of platinum complexes and cyclometals, which is applied in the field of biological analysis, can solve the problems of difficult control of staining conditions in silver staining, poor sensitivity of Coomassie brilliant blue, and high operating level requirements, and achieve high staining sensitivity, low cytotoxicity, and sensitivity high effect

Inactive Publication Date: 2019-07-30
SUZHOU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The dyes used in colorimetry include silver nitrate and Coomassie brilliant blue, etc., but silver nitrate and protein staining are combined through irreversible chemical bonds, which is not conducive to the subsequent analysis of proteins. Higher levels required; Coomassie Brilliant Blue less sensitive
The dyes used in the fluorescence method include two categories: small molecules and metal complexes. Metal complex luminescent dyes occupy an important position in the protein stain market due to their high stability and high luminous efficiency; however, the existing metal complexes Poor stability, high cytotoxicity, low staining sensitivity, about a few hundred nanograms, and complicated washing steps

Method used

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  • A kind of cyclometal platinum complex and its preparation method and its application as protein staining agent
  • A kind of cyclometal platinum complex and its preparation method and its application as protein staining agent
  • A kind of cyclometal platinum complex and its preparation method and its application as protein staining agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Synthesis of Cyclometallic Platinum Complexes

[0039]

[0040] First, potassium tetrachloroplatinate and C^N bidentate ligand were reacted in ethylene glycol ether solvent for 24 hours; then O^O bidentate ligand compound was added to continue the reaction for 24 hours, the solvent was spin-dried, and column chromatography The ring metal platinum compound was isolated. Yield: 56%. Mass spectrum: experimental value [M+Na] + 737.1409, theoretical value [M+Na] + 737.1408.

[0041] By replacing C^N bidentate ligands and N^N bidentate ligand compounds, cyclometal platinum complexes with multiple structures can be obtained.

[0042] The following examples use cyclometal platinum complexes for protein staining and gel testing. The process is as follows:

[0043] After the commercialized protein samples were subjected to SDS-PAGE, they were fixed in 30% ethanol and 10% acetic acid; the gel was rinsed in 20% ethanol for 3 times, 30 minutes each time; The subs...

Embodiment 2

[0045] The structural formula of the protein stain used is:

[0046]

[0047] The fluorescence quantum efficiency of the compound is 37%, with figure 1 Its fluorescence picture; dilute the protein stain luminescent platinum complex from the mother solution (with DMSO as solvent) to 1 micromol with pure water for staining.

[0048] Taking HeLa cells (HeLa), MCF-7, and A549 cancer cell organelles as objects, the 24-hour cytotoxicity IC50 value of the platinum compound in this example is 150 micromolar, which proves that the cytotoxicity of this platinum complex is very small.

Embodiment 3

[0050] The structural formula of the protein stain used is:

[0051]

[0052] The fluorescence quantum efficiency of the compound is 46%, with figure 2 Its fluorescence picture; dilute the protein stain luminescent platinum complex from the mother solution (with DMSO as solvent) to 1 micromol with pure water for staining.

[0053] Taking HeLa cells (HeLa), MCF-7, and A549 cancer cell organelles as objects, the 24-hour cytotoxicity IC50 value of the platinum compound in this example is 150 micromolar, which proves that the cytotoxicity of this platinum complex is very small.

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Abstract

The invention discloses a ring metal platinum complex as well as a preparation method and an application thereof as a protein coloring agent. The ring metal platinum complex comprises a C-N di-tooth ligand and / or an O-O di-tooth ligand and is a light-emitting metal complex. The preparation method is simple and the ring metal platinum complex has a relatively high quantum efficiency. As the protein coloring agent, the ring metal platinum complex is high in coloring sensitivity which can reach an ng level. The rinsing step is cancelled, so that the time is shortened; the defects in the prior art that rinsing is needed and the sensitivity is low are solved, and the ring metal platinum complex disclosed by the invention is simple in preparation method, high in stability and relatively low in cytotoxicity.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and in particular relates to a cyclometal platinum complex, a preparation method thereof and an application as a protein stain. Background technique [0002] As an important technique, electrophoresis is widely used in biologically related fields such as protein analysis and drug development. By means of electrophoresis, proteins can be separated into different bands on sodium dodecylsulfonate-polyacrylamide gel plates (SDS-PAGE) according to different molecular weights. The visualization of these protein bands is the premise and basis for subsequent protein analysis. [0003] Protein visualization methods currently include colorimetric and fluorescent methods. The dyes used in colorimetry include silver nitrate and Coomassie brilliant blue, etc., but silver nitrate and protein staining are combined through irreversible chemical bonds, which is not conducive to the subsequent analys...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07F15/00C09K11/06G01N1/30
CPCC07F15/0086C09K11/06C09K2211/185G01N1/30
Inventor 周宇扬
Owner SUZHOU UNIV OF SCI & TECH
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