Bio-Synthesis offer custom ascites monoclonal antibodies (mAb) from BALB/c-derived cell lines by utilizing a multi-phase production process with an extensive quality assurance program. These monoclonal antibodies are generated by injecting the hybridoma cell line into the primed peritoneal cavity of BALB/c mice and harvesting the resulting fluid that contains antibodies secreted by the hybridoma. Customers will supply cell line(s) of interest to Bio-Synthesis for the production of mouse ascites or we can provide cell line produced through our monoclonal antibody development program. The custom ascites production service is based on number of mice used to generate the quantity of ascites needed by our customer. Average ascites yield for Balb/c is 3ml. Actual yield will vary for each cell line.
To obtain further information regarding our ascites monoclonal antibody production services, please contact our National Customer Services Center at 800.227.0627 or contact us online. BSI supplies an Ascites Production Order Form with each inquiry to assist in meeting customer specifications.
We offer discount for large orders, please contact our sales and marketing department for further details.
BSI requires two hybridoma samples for ascite production. The first hybridoma sample is used to screen the hybridomas for mycoplasma contamination. The second hybridoma sample is used to be cultured and injected into the production mice.
Hybridomas with more than 90% viability that are rapidly growing and dividing (indicative of the log phase), should be used for cryo-preservation (freezing). The hybridomas should be grown in an antibiotic-free medium (DMEM or RPMI with 10% FBS) for a minimum of two passages. Working in a sterile environment, remove the hybridoma cells from the flask, then transfer to a centrifuge tube. After counting the cells, determine the necessary volume to have 5 x 106 hybridomas per cryo-vial. Centrifuge the hybridoma cells at 100 g for 10 minutes. Remove the media carefully and re-suspend the hybridoma cells in freezing media (90% fetal bovine serum (FBS) and 10% DMSO) in order to have 5 x 106 hybridoma cells per ml in each cryo-vial. Note, the hybridoma cells should be frozen slowly. In order to do this, the vials must be enclosed within two polystyrene tube racks by placing the cryo-vials in one polystyrene tube rack. The second rack is then inverted and placed over the tops of the vials. The rack halves are then taped together and left overnight in a -80°C freezer (a special cryogenic freezing container can also be used). The next day, transfer the cryo-vials into a liquid nitrogen tank or ship them using next-day delivery with a minimum of 10 lbs of dry ice included in the shipment. When working with liquid nitrogen and frozen cryo-vials, use gloves and a face mask as protecting gear. It is recommended that the duplicate samples be prepared from the same hybridoma culture to be tested for mycoplasma and preparation of ascites fluid.
To maximize mycoplasma detection, it is recommended that the hybridomas be grown for a minimum of two passages in an antibiotic-free medium. This is crucial because antibiotics usually suppress, not eliminate, mycoplasma growth. The hybridomas should also be harvested when they are growing rapidly in the log phase.(Note: It is not recommended to use Trystin-EDTA for harvesting.) For optimal results, 1,000 to 10,000 cells should be frozen under sterile conditions.