|Anti-TFPI Stable Cell Line
- Cell Line Description
- This gene encodes a protease inhibitor that regulates the tissue factor (TF)-dependent pathway of blood coagulation. The coagulation process initiates with the formation of a factor VIIa-TF complex, which proteolytically activates additional proteases (factors IX and X) and ultimately leads to the formation of a fibrin clot. The product of this gene inhibits the activated factor X and VIIa-TF proteases in an autoregulatory loop. The encoded protein is glycosylated and predominantly found in the vascular endothelium and plasma in both free forms and complexed with plasma lipoproteins. Several alternatively spliced transcript variants of this gene have been described, but the full-length nature of some of these variants has not been confirmed.
- 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
- HPLC > 98%, One strip for SDS-PAGE.
- Humanized IgG4 - kappa
- Product Storage
- It should be stored in liquid or vapor phase nitrogen. Reconstituted antibody aliquots should avoid repeated freeze-thaw cycles.
- Involvement in Disease
- Concizumab (mAb 2021) is a monoclonal IgG4 antibody (mAb) that binds to the Kunitz-type protease inhibitor (KPI) 2 domain of TFPI thereby blocking the interaction of this domain with the active site of FXa. It promotes coagulation by neutralising the inhibitory function of TFPI and may provide a subcutaneous prophylaxis option for patients with haemophilia.
- Concizumab is a monoclonal antibody (mAb) against Tissue Factor Pathway Inhibitor (TFPI).
- TFPI; tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor); LACI; tissue factor pathway inhibitor; EPI; extrinsic pathway inhibitor; TFI; TFPI1; anti-convertin;
- Complement and Coagulation Cascades, organism-specific biosystem; Complement and coagulation cascades, organism-specific biosystem; Complement and coagulation cascades, conserved biosystem; Extrinsic Pathway, organism-specific biosystem; Formation of Fibrin Clot (Clotting Cascade), organism-specific biosystem; Hemostasis, organism-specific biosystem; Syndecan-4-mediated signaling events, organism-specific biosystem;
- endopeptidase inhibitor activity; serine-type endopeptidase inhibitor activity;
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