Protein protectant

A technology of protein protectant and protectant, applied in the biological field, can solve the problem of long-term storage of Hsp90α protein that has not been seen

Active Publication Date: 2016-12-07
YANTAI PROTGEN BIOTECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the different properties of proteins, it is difficult to establish a general protective agent formula for different proteins, so the development of bet

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0121] Example 1 Effect of adding a single component protective agent on the stability of Hsp90α protein

[0122] Objective: To screen new Hsp90α protein protectors, and to determine the concentration range and conditions for their use.

[0123] Reagents: Phosphate, Tween 20 and tea polyphenols were purchased from Sinopharm Chemical Reagent Co., Ltd., bovine serum albumin was purchased from Sigma Company, heparin sodium was purchased from Xinjing Biotechnology, and cyclodextrin was purchased from Beijing Aoboxing Biotechnology Co., Ltd. company. The Hsp90α finished kit was purchased from Yantai Proji Biotechnology Development Co., Ltd., batch number 201602002, 201604001, and the calibrator of the kit was Hsp90α freeze-dried product.

[0124] method:

[0125] 1. Prepare the basic solution, use purified water, which contains 0.318% phosphate (sodium dihydrogen phosphate), 0.5% bovine serum albumin and 0.2% Tween 20, adjust the pH to pH6.5, this solution is the basic of the lat...

Embodiment 2

[0151] Example 2 Effects of Combined Use of Multiple Protective Agents on the Stability of Hsp90α Protein

[0152] Objective: To develop a new combination formula of Hsp90α protein protector, and to determine its concentration range and conditions.

[0153] Reagents: Phosphate, Tween 20, MgCl 2 , tea polyphenols and K 2 SO 4 Purchased from Sinopharm Chemical Reagent Co., Ltd., bovine serum albumin and AMP were purchased from sigma, heparin sodium was purchased from Xinjing Biotechnology, PVP40 was purchased from Xinjing Biotechnology, and cyclodextrin was purchased from Beijing Aoboxing Biotechnology Co., Ltd. The Hsp90α finished kit was purchased from Yantai Proji Biotechnology Development Co., Ltd., batch number 201602002, 201604001, and the calibrator of the kit was Hsp90α freeze-dried product.

[0154] method:

[0155] The basic experimental method and steps are the same as those in Example 1, and in this example, a combination of multiple protective agents is used.

...

Embodiment 3

[0183] Example 3 Using the most effective protective agent combination formula to detect the performance index of the Hsp90α kit

[0184] Objective: To test the effect of the combination formulation of the most effective protective agent on the detection performance of Hsp90α protein and the kit through 12 days of accelerated experiment at 37°C.

[0185] Method: The detection was performed according to the operation steps in Example 1, wherein the accelerated Hsp90α freeze-dried product was used as the calibration product curve, and the quality control product was added at the same time to test the performance of the kit.

[0186] Results: The Hsp90α freeze-dried calibrator using the most effective combination formula is still stable after 12 days of acceleration, and the results of various performance indicators of the kit are shown in Table 7 below:

[0187] Table 7 The structure of each performance index of the kit

[0188]

[0189] Among them, the lowest detection limi...

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Abstract

The invention provides a series of protein protective solutions which are capable of effectively protecting the activity of Hsp90 (Heat Shock Protein90) alpha protein and increasing the stability of the Hsp90alpha protein and are invariant under the concentration in nanogram level. On the basis of a traditional protein protectant, other protectant components are added in the protein protective solutions, and the other protectant components are selected from AMP (Adenosine Monophosphate), heparin sodium, MgCl2, PVP40 (Polyvinylpyrrolidone 40), tea polyphenol, cyclodextrin and K2SO4.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein protective agent for tumor marker Hsp90α. Background technique [0002] Heat shock protein 90α (Heat shock protein 90α, Hsp90α) is one of the most abundant chaperone proteins in the cytoplasm of eukaryotic cells, accounting for about 1-2% of the total protein content in normal cells. In tumor cells, the content of Hsp90α can be as high as 2-7% of the total amount of intracellular proteins (Whitesell, L. and S.L.Lindquist., 2005, Nat Rev Cancer. 5 (10), 761-772), which can improve tumorigenesis and Both the stability and the assistance of the function of related protein factors during development are necessary. Studies have shown that Hsp90α is highly expressed on the surface of many tumor cells, including small cell carcinoma, melanoma, and liver cancer cell lines (Ferrarini et al., 1992, Int. J. Cancer. 51, 613-19). Another article reported that Hsp90α secreted into the ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/574G01N33/68G01N2333/46
Inventor 崔大伟刘艳李米川徐红
Owner YANTAI PROTGEN BIOTECH DEV
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