|Anti-HBV Stable Cell Line
- Cell Line Description
- HBV (HBV) infection induces a disease state which manifests itself in a variety of ways, characterized by the extent of liver damage, inflammation and viral persistence. HBV infection is also associated with a 100 fold increased risk of hepatocellular carcinoma and currently infects over 250 million people worldwide. HBV has a partially double stranded 3.2 kilobase DNA genome which contains four open reading frames. One of these encodes a 154 amino acid protein called the HBx protein. HBx has been shown to be a transcriptional transactivator of both viral and cellular promoters. Lacking a DNA binding domain and nuclear localization signal, HBx is believed to exert transcriptional activity through protein protein interaction.
- Growth Properties
- Complete growth medium: Serum-free Medium
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
- Starting Cells From Frozen Cell Stock
- 1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37°C water bath for 2 minutes until nearly (80%) thawed.
Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.
2. Clean the outside of the vial with 70% ethanol before opening.
3. Remove the cells from the vial and add slowly into a 15ml conical tube containing 10 ml pre-warmed media.
4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.
5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.
6. Place the cells in the 37°C incubator with 5% CO2.
7. Allow incubation for 3-4 days to reach confluence. The cells will re-attach to the surface over a period of several days in culture at 37°C.
- 1. Centrifuge for 5 minutes 300 xg and discard culture medium.
2. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
3. Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
- Mycoplasma Status: Negative (MycoAlert Kit)
- Freeze Medium
- Complete growth medium 90%; DMSO, 10%
- Safety Considerations
- The following safety precautions should be observed.
1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.
2. No eating, drinking or smoking while handling the stable line.
3. Wash hands after handling the stable line and before leaving the lab.
4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.
5. All waste should be considered hazardous.
6. Dispose of all liquid waste after each experiment and treat with bleach.
- Product Purity
- >98.0% as determined by Analysis by RP-HPLC & analysis by SDS-PAGE.
- Product Molecular Mass
- Approximately 150 kDa
- Product Storage
- It should be stored in liquid or vapor phase nitrogen. Reconstituted antibody aliquots should avoid repeated freeze-thaw cycles.
- Involvement in Disease
- Libivirumab is a human monoclonal antibody directed against the hepatitis B virus
- This is a human monoclonal antibody directed against the hepatitis B virus.
- HBsAg; Hep B surface antigen; Hepatitis B Virus major surface antigen; Hepatitis B virus S antigen; Large envelope protein; Large surface protein; LHB; Major surface antigen
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