Monoclonal FAQ’s

Q: What is the antibody yield from ascites fluid?

A: The yield of monoclonal antibody in ascites fluid depends on two major factors. First, the inherent ability of the hybridoma cell line to synthesize and secrete monoclonal antibody. This inherent monoclonal antibody production rate may be improved with molecular biology intervention. Second, the health, clonality, and environment in which the hybridoma cell line is synthesizing and secreting monoclonal antibody contribute to the productivity of the cell line. The full inherent potential of the cell line to produce monoclonal antibody may be compromised if the culture viability is poor, if the cell line is allowed to exhaust critical media components or if the clonality of the cell line is low.

SDIX’s mean antibody yield from ascites production using hundreds of cell lines over the last 2 years is 18.9 mgs of antibody per mouse. The mean has a wide variance due to inherent different among cell lines to synthesize and secrete monoclonal antibody. Some cell lines have produced less than 1 mg of antibody per mouse while others have produced over 50 mgs of antibody per mouse.

Q: What is the antibody yield from roller bottle supernatant?

A: The yield of monoclonal antibody also depends on the inherent ability of the cell line to synthesize and secrete monoclonal antibody and the health, clonality and environment in which the hybridoma cells are synthesizing and secreting monoclonal antibody. Changes in media formulation may improve the antibody yield. SDIX means antibody yield from roller bottle supernatant is 35 mgs per liter. This mean has a large variance due to inherent differences among cell lines to synthesize and secrete monoclonal antibody and due to the improvements in yield realized by manipulating the media formulation. Some cell lines produce less than 10 mgs of antibody per liter while others produce over 50 mgs per liter.

Q: Which method should I use for monoclonal antibody production, ascites or roller bottles?

A: Three major factors are used to determine the production mode: economics, ethical considerations, and the desire to define the environment in which the hybridoma cell line produces antibody.

Economics: For most cell lines, the cost of antibody produced in ascites is less than the cost of antibody produced using roller bottles. This difference becomes very important when tens or hundreds of grams of monoclonal antibody are produced.

Ethical considerations: Many individuals and many organizations choose to avoid the use of animals when a suitable alternative to the use of animals is available. All of SDIX’s animal facilities are AAALAC-accredited.

Defining the environment in which the hybridoma cell line produce antibody: Ascites fluid is a complex mixture of many mouse proteins and other biomolecules. These molecules are present in the ascites product. The identities and concentrations of all of these proteins and biochemicals is not known. These molecules are usually removed during purification of monoclonal antibody from ascites fluid. To remove the uncertainty of the influence of the these proteins and biomolecules on the outcome of antibody purification, production of monoclonal antibody is roller bottles is available. The environment in which the hybridoma cell line makes antibody (the media formulation) is easily defined, especially when serum-free media formulations are used.

Q: Why use SCID mice for ascites production?

A: SCID is an acronym for Severe Combined ImmunoDeficient mice. These mice lack the ability to synthesize and secrete endogenous antibodies. When SCID mice are used for ascites production, the only antibody present in the ascites fluid is the monoclonal antibody produced by the hybridoma cell line used for ascites production.

Q: What do need to get started?

A: To produce antibody in ascites fluid or in roller bottles, we require frozen cryovials of the hybridoma cell line and a completed Project Submission Form. This form captures your antibody production experience with the cell line (if any) and information required for successful antibody production (such as the media formulation).

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