Production of Monoclonal Antibody in Mouse Ascitic Fluid

• Culture of hybridoma for 15 days
• Freezing of aliquots of hybridoma cells: 2-8.106 cells per vial (3 vials)

 

Covalab freezes aliquots of hybridoma cells in vials only on the client's request. The client must indicate on his order whether Covalab must return the vials at the end of production or whether the vials must be stored at Covalab. If no mention of this is made on the order, no vials will be prepared.

 

All hybridomas must be accompanied by a certificate, testifying to the absence of mycoplasma.

The client reserves the right not to order cell culture. Under such circunstances Covalab does not take responsibility for the production of antibodies which do not meet its quality control procedure in cell culture.

• If Covalab is performing the hybridoma culture, the cells must be frozen and the vials shipped in sufficient dry ice to last for 48h maximun. We prefer to receive 2 vials.
• If cell culture is not ordered, cells in the exponential growth phase must be shipped in culture medium at room temperature within the shortest delay possible to reduce cell death to a minimum.
• On receipt of the cells, a viability assay is performed so that the required number of viable cells is injected to each mouse. In view of this, the number of cells sent must be proportional to the number of mice ordered. At the end of the injection any excess cells are destroyed.

• Pristaned female mice (9 to 13 weeks old)
• Intraperitoneal inoculation of hybridomas cells: 1.106 cells/mouse 3 weeks after receiving pristane
• Stabulation of animals (35 days from injection date)
• Collection of ascitic fluid
• Conservation of ascitic fluid with 0.02% sodium azide

Remarks:

• The service is performed for a minimum order of 4 mice.
• 1 mouse produces around 2-4 ml of ascitic fluid
• The concentration of mouse immunoglobulins in an ascitic fluid can vary from 1 to 5 mg/ml and of this amount, 80 to 90% corresponds to specific antibody. It should be noted that the yield of antibody by hybridomas is dependent on their capacity to secrete antibody. This varies from 1 hybridoma to the next.
• Any control of the immunoreactivity is performed by Covalab

Purification of antibodies
Labelling of antibodies

At the end of the contract, Covalab proposes to store the hybridomas in vials frozen in liquid nitrogen. Minimum storage period: 1 month.

Please contact us