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