Process for the purification of Fc-fusion proteins
A technology of fusion protein and protein, which is applied in the field of protein purification, can solve the problems of not proposing systematic removal of specific order, not being able to meet the requirements of sufficient purification, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0306] Example 1 : capture step: protein A affinity purification
[0307] The starting material is the clarified collection of CHO cell clones expressing TACI-Fc cultured under serum-free conditions, and stored frozen until use.
[0308] In MabSelect Xtra according to the following scheme TM This capture step was performed on a column (GE Healthcare 17-5269-03) with a bed height of 17 cm. All manipulations were performed at room temperature, except that the loading solution was kept below 15 °C. Record the UV signal at 280 nm.
[0309] clean
[0310] The column was cleaned with at least 3BV of 0.1 M acetic acid + 20% ethanol at a counterflow rate of 250 cm / h. Stop flow for 1 hour.
[0311] washing steps
[0312] The column was washed with at least 2BV of RO water at a counterflow rate of 250 cm / h.
[0313] balance
[0314] Equilibrate with at least 5BV of 25mM sodium phosphate + 150mM NaCl solution pH7.0 (unit conductivity and pH parameters are within a specific rang...
Embodiment 2
[0337] Example 2 : Cation Exchange Chromatography
[0338] The eluate from the protein A capture step was dialyzed against the appropriate loading buffer and used as starting material for cation exchange chromatography.
[0339] This step uses Fractogel EMD SO with a bed height of 10cm 3 - Column (Merck 1.16882.0010). Fractogel EMD SO with a bed height of 15 cm can also be used 3 - column. The latter's dynamic capacity and flow rate may require modification, which is within the routine knowledge of those skilled in the art.
[0340] All operations were performed at room temperature and the flow rate was kept constant at 150 cm / h. The 280nm UV signal was recorded for all time periods.
[0341] washing steps
[0342] The column was washed with at least 1 BV of WRO (water for reverse osmosis).
[0343] clean
[0344] Then, clean the column with at least 3BV of 0.5M NaOH + 1.5M NaCl in an upflow fashion.
[0345] rinse
[0346] Rinse the column with at least 4BV of W...
Embodiment 3
[0381] Example 3 : Anion exchange chromatography
[0382] The starting material used in this purification step is Fractogel SO 3 - (See Example 2) The eluate from the cation exchange step is dialyzed or diluted with the appropriate loading buffer.
[0383] This step of anion exchange chromatography was performed on a SOURCE 30Q chromatography column (GE Healthcare 17-1275-01) with a bed height of 10 cm. In this step, a SOURCE 30Q chromatographic column with a column bed height of 15 cm can also be used. The latter dynamic capacity and flow rate may require adaptation, which is within the routine knowledge of those skilled in the art.
[0384] All operations were carried out at room temperature, and the signal of 280nmUV was recorded. This step is performed at a flow rate of 150 or 200 cm / h.
[0385] rinse
[0386] Wash the column with at least 1BV of RO water at a flow rate of 150cm / h.
[0387] clean
[0388] The column was then cleaned with at least 3BV of 0.5M NaOH...
PUM
| Property | Measurement | Unit |
|---|---|---|
| pore size | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More 
PatSnap Eureka turns technology decisions into work you can execute. Powered by our Innovation Knowledge Graph, it runs expert workflows across engineering, life sciences, materials and intellectual property. Get your review-ready output in minutes.
