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1-90401: Custom Monoclonal Antibody Production |
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Introduction: |
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This package is designed for
generating custom monoclonal
antibodies against a synthesized
peptide or recombinant protein
segments. |
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What is included with this
package: |
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Phases |
Details |
Cost |
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Phase 1:
Immunization |
Immunization of Balb/c
mice ( up to 5 animals) with
antigen to stimulate
antibody production. You may
choose to follow our
standard immunization
protocol, or work with our
staff to use your own
protocol. We typically
inject mouse every two weeks
for 6 weeks before taking a
test bleed. The test bleed
serum samples are analyzed
for antibody production
using ELISA. If desired,
clients can request up to
0.5 ml serum sample/per
mouse for their own
additional characterization.
ELISA will be used to
determine the highest titer
mice to be used for cell
fusion. Immunization may
continue longer if response
is not optimal.
Antigen requirement:
0·5
mg of purified recombinant
protein or intact protein
will be needed for
immunization.
With peptide synthesis
request: Custom Peptide
synthesis up to 15 amino
acids (additional amino acid
for $15 per residue), HPLC
purification up to 90%
purity, peptide KLH
conjugation for
immunization, peptide BSA
conjugation for screen and
10 mg free peptide will be
provided for client.
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8-10 weeks ($1,000
without peptide
synthesis)
or
($1,500 with peptide
synthesis) |
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Phase 2:
Fusion |
Fusion of spleen cells with
myeloma cell line. The fused
cells are then placed in
several 96-well culture
plates. Viable colonies
appear as early as 3-5 days
after fusion and are kept
healthy with a fresh supply
of nutrient media as
necessary. |
2-6 weeks $1,500 |
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Phase 3:
Screening |
When hybridoma colonies are
large enough to screen,
screening of all wells
containing fusion products
is done with ELISA assay.
All positive wells will be
expanded and frozen for
parental hybridomas. The
number of positives depends
on the nature of the
antigen, and can range from
a few hybridomas to a dozen
or more. Additional hybrids
can be frozen for further
studies. |
2-6 weeks $1,500 |
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Phase 4:
Stabilization and Expansion |
Expansion of clones
After positive hybridomas
are selected for further
development, they will be
cloned using the limited
dilution method and screened
by ELISA. 5 clones with the
desired specificity will be
expanded, and hybridoma
medium (50 - 100 ml) will be
collected for further
studies |
2-4 weeks $1,000 |
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Phase 5:
Isotyping of clones and
Ascites Production |
Isotypes of all selected
clones will be determined
and production of ascites
from 5 mice if needed. |
3-5 weeks $1,000
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Total |
Complete custom
monoclonal production |
17-29 weeks ($5,999) |
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Additional Options |
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Purification |
The secreted antibody
will be purified by Protein
G affinity chromatography
and send to the customer.
Approximate yield may be
20-30 mg.
Antibodies are eluted using
both pH11 and pH2.5 buffers
and neutralized immediately
followed by dialysis in PBS. |
1 week
$500/per project |
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