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Phase I
*Immunization of two New Zealand rabbits.
*Evaluation of antibody response by ELISA,
Western blot, and/or IHC.
Approximately 3-4 mg protein immunogen or 3
mg conjugated peptide immunogen and 1 mg
free peptide are required.
Estimated time: 6~8 weeks
Price: $1,500
Phase II
*Separation of ASC
(antibody-screening plasma cell).
*RT-PCR of V-heavy and V-light variable
regions.
*Construct E. coli expression clones with
both heavy and light chains.
*Screening of clones in ELISA.
*V-heavy and V-light variable regions are
cloned into IgG1 or IgK expression vector.
*The vectors are co-transfected in HEK-293,
and the supernatant from post-transfection
is used for antibody screening by ELISA.
*1ml of supernatant of 293 cell transfection
from up to 10 clones will be sent to the
customer for evaluation. Customer can use
HRP-labeled anti-rabbit as secondary
antibody for the assay.
*Sequencing of V-heavy and V-light variable
regions of clone with positive antibody
response.
Estimated time: 4~6 weeks
Price: $10,000
Phase III
*Scale-up of transient transfection to make 5
to 10mg of full length antibody that will be
purified from HEK-293 cell culture
supernatant using protein A/G column and
delivered to our client.
Estimated time: 2 to 4 weeks
Price: $3,500
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