Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Detection method of protein content in drug gel

A technology of protein content and detection method, which is applied in the field of detection of protein content in pharmaceutical gels, can solve the problems of inaccurate quantification, difficulty in controlling the release amount, and large differences in results, etc., and achieves easy to master operation, reasonable design, and repeatable strong effect

Inactive Publication Date: 2014-05-14
BEIJING SL PHARMA +2
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The disadvantage of the above method is: the colloidal structure of the gel matrix carbomer is stable, and the drug protein in the gel cannot be completely released only by blowing, and the amount of release is difficult to control, which then leads to inaccurate quantification and poor repeatability. Limited use in research
However, in practice, the measured results vary widely
After analysis, the reason for the inaccurate determination is that the existing detection method cannot completely release rh-bFGF from the gel

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Detection method of protein content in drug gel
  • Detection method of protein content in drug gel
  • Detection method of protein content in drug gel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Taking the detection of the rh-bFGF content in the recombinant human basic fibroblast growth factor (rh-bFGF) gel as an example.

[0035] bFGF gel is the latest new drug product of Beijing Shuanglu Pharmaceutical Co., Ltd. for the treatment of tissue wound healing. Each rh-bFGF gel for external use contains 5g rh-bFGF gel, and each 1g contains 5000IU rh-bFGF.

[0036] 1. The process prescription is:

[0037]

[0038] 2. Preparation process:

[0039] According to the calculated amount, take glycine and ethyl 4-hydroxybenzoate by weighing respectively and put into (two-thirds of the total amount) water for injection to prepare the aqueous solution of glycine and ethyl 4-hydroxybenzoate. Heat to dissolve and stir well. Take a quarter of it and autoclave it for later use (liquid A).

[0040] Weigh the carbomer according to the calculated amount and dissolve it in the above aqueous solution to make a carbomer gel, which is autoclaved for future use (liquid B).

[0041]...

Embodiment 2

[0053] Determination of the content of drug protein rh-bFGF in bFGF gel

[0054] 1. Determination of rh-bFGF microplate reader method detection standard working curve

[0055] Accurately measure 10mL of rh-bFGF solution with a concentration of 12.5mg / ml, add double distilled water to make the volume to 250mL, dilute the rh-bFGF standard to 50μg / ml, and accurately measure 0, 10, 20, 40, 60, 80, 100 and 120 μl were sucked into 1.5ml eppendorf tubes, and the gel blank was added to 800 μl, and 200 μl Bradford working solution was added to each tube (preparation of Bradford working solution: take 25ml 95% ethanol, 50ml 85% phosphoric acid, 50mg Coomassie brilliant blue G- 250 to mix evenly, add distilled water to dilute to 500ml), develop color for 5 minutes, pipette 250μl into a 96-well plate, and scan the absorbance value at 540nm with a microplate reader.

[0056] Take the rh-bFGF standard substance concentration as the ordinate and the light absorption value at 540 nm as the a...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a detection method of drug protein content in drug gel, and is characterized in that when drug protein is released from gel, the gel is diluted or not diluted at first; a solution containing calcium ions is added as a releaser, wherein the calcium ion adding concentration range is 0.015 mol / L-0.15 mol / L. The gel is precipitated rapidly, and the drug protein is released completely from the gel, which enables more accurate determination of the drug protein content in the drug, and the biological activity titer. The method is suitable for the detection of drug protein content in gel drugs with carbomer used as a gel matrix, such as the detection of rh-bFGF content in recombinant human basic fibroblast growth factor gel. The method is simple, accurate, and strong in repeatability.

Description

technical field [0001] The invention relates to a method for detecting protein content in drug gels, which is suitable for detecting drug protein content in gel drugs with carbomer as the gel matrix, and is especially suitable for detecting the growth of recombinant human basic fibroblasts Factor (hereinafter referred to as rh-bFGF) content of rh-bFGF in the gel. Background technique [0002] Existing drug gels, especially gel preparations of protein and polypeptide drugs, mostly use carbomer as the gel matrix. The content of drug protein in some of these gel medicines is very small, and some preparations only contain more than ten micrograms of drug protein per gram of gel. Since it is difficult to release all the trace drug proteins bound in the carbomer hydrogel of the three-dimensional network structure, the content and biological activity of drug proteins in the drug cannot be accurately determined when detecting the drug content of this type of gel. [0003] At prese...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 徐明波吴彦卓陈献华徐霞王俊玲杨仲璠秦富景白福英倪娜赵紫岭刘琼
Owner BEIJING SL PHARMA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com