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).