A kind of preparation method of feiba with human plasma cohn component iii as raw material
A technology for human plasma and raw materials, which is applied in the preparation methods of peptides, chemical instruments and methods, thrombomodulin, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0076] Preparation of FEIBA Forming Raw Materials from Component III
[0077] 1. Weigh 3 kg of the plasma Cohn fraction III precipitate, and put it into 12 kg of reconstitution buffer at a ratio of 1:4 (the buffer is 0.01M sodium citrate + 0.1M sodium chloride aqueous solution, pH7.5 ). Stir at a temperature of 25±1°C, adjust the pH to 7.0-7.5 with 1.0M aqueous sodium bicarbonate solution, and continue stirring until the precipitate is completely dissolved.
[0078] 2. Add PEG4000 aqueous solution (40wt%) to the above component III complex solution to a final concentration of 5%, and lower the temperature to 1-4°C, and continue stirring for 30 minutes. At this time, the pH of the solution is about 7.0-7.4.
[0079] 3. The complex solution of component III is centrifuged at low temperature to separate the precipitate and supernatant. Centrifuge at a speed of about 10000g, and centrifuge at a temperature of 1-4°C for 10 minutes. The supernatant after centrifugation was collec...
Embodiment 2
[0085] 1. Immerse the A50 gel that has been treated with acid and alkali and sterilized at 121°C in an aqueous solution of pH 7.5, 0.075M sodium chloride + 0.1M sodium citrate to balance. Measure the well-balanced A50 gel in a ratio of 1g / L, and put it into the 2Kg feed solution prepared in Example 1.
[0086] 2. After stirring and adsorbing for 1 hour, collect the gel by filtration, wash with pH 7.5, 0.1M sodium chloride + 0.01M sodium citrate aqueous solution for three consecutive times, and discard the filtrate. Finally, it was eluted with pH 7.5, 1M sodium chloride+0.01M sodium citrate aqueous solution, and the eluate was collected. This step operation is carried out at room temperature.
[0087] 3. Take 40ml of the eluate, use a 10KD ultrafiltration centrifuge tube, carry out ultrafiltration and concentration at 5000g, 4°C, and replace the liquid to 9.3ml. The replacement buffer is 0.05M Tris-Hcl, 0.15M NaCl, pH7.0, A280 is 54.8.
[0088] 4. Put the concentrated soluti...
Embodiment 3
[0093] 1. Take 2kg of the S / D feed solution prepared in Example 1, put the pretreated A50 gel into it at a ratio of 1g / L, stir at room temperature for 1 hour, and collect the gel by filtration.
[0094] 2. Treat the gel in step 1 with 0.1mol / L NH at pH 7.6 4 HCO 3 Wash about 200ml with aqueous solution, 0.2mol / L NH at pH7.6 4 HCO 3 Wash about 150 ml with aqueous solution, and about 200 ml with 0.15 mol / L NaCl aqueous solution, and discard the filtrate. Finally, the protein adsorbed by the gel was eluted with 1 mol / L NaCl aqueous solution, and about 150 ml of the eluate was collected.
[0095] 3. Under the condition of 4°C, use the replacement buffer solution (pH7.0, 0.05M Tris-HCl+0.15M NaCl aqueous solution) to carry out ultrafiltration concentration and exchange, and the pore size of the ultrafiltration membrane is 10KD. The final 150ml eluate was concentrated to about 30ml.
[0096] 4. Take a part of the concentrated solution and place it in an environment of 12°C. Aft...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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