Polyclonal antibodies by
genetic immunization
Although the use of monoclonal antibodies holds promise for
the detection of the respective antigen with high sensitivity,
for some applications polyclonal antibodies are superior due
to faster availability and broader detection of antigen by
the multivalent but monospecific nature of antisera.
ViroLabs offers the production of antisera by genetic immunization
of mice, rats and rabbits.
Please provide the gene encoding the protein antigen against
which the antibodies shall be generated cloned into any plasmid
vector. Several microgram of the vector in an eppendorf tube
or simply dropped on a small peace of filter paper are sufficient.
Basic study design for rabbit immunization:
Day 0 Pre-immunization blood draw (2ml serum) and genetic
immunization
28 Genetic immunization
56 Genetic immunization (test serum possible two weeks later)
84 final genetic immunization (optional)
98 final bleeding (at least 30-40 ml serum)
Additional DNA or protein immunization and bleedings are
possible. The quantity of DNA used per immunization event
in the basic study design is 1 mg. We recommend to immunize
two animals with each construct.
Unless otherwise specified, antisera will be aliquoted as
one tube per animal and shipped in a chilled state.
Monoclonal antibodies by genetic immunization
Monoclonal antibodies from genetic immunization approaches
are often superior compared to monoclonals derived from conventional
protein or peptide based immunizations in terms of affinity
and specificity.
ViroLabs offers the production of monoclonal antibodies by
genetic immunization of mice and rats. Please provide the
gene encoding the protein antigen towards which the antibodies
shall be generated cloned into any plasmid vector. Several
microgram of the vector in an eppendorf tube or simply dropped
on a small peace of filter paper are sufficient.
The delivery time of the antibodies depends mainly on the
immunogenicity of the respective antigen and the amount of
subcloning. Usually it takes between 6 and 7 months.
Animals are immunized until a satisfactory titer is detected.
A standard schedule is given below. Modifications are possible.
Basic study design for mouse immunization:
Day 0 Pre-immunization blood draw and genetic immunization
28 Genetic immunization
56 Genetic immunization (titer testing)
84 Final immunization (titer testing)
98 Fusion
112 Screening
133 Subcloning
Additional DNA or protein immunization and bleedings are
possible. The quantity of DNA used per immunization event
in the basic study design is 100 µg. The fusion of B
cells and myeloma cells is done by the PEG method. Hybridoma
cells are screened for antibody production and the best candidates
are cloned. Subcloning is done by repeated limiting dilution
technique to obtain a monoclonal hybridoma cell culture.
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