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We will link two approved anti-tumor antibiotics to your purified monoclonal antibody using our proprietary conjugation method. The anti-tumor agent will remain bound to the monoclonal antibody and inactive but will be released and activated by reduction once the immunocomplex binds to the cell surface antigen and internalized.

1. Pilot Study

Material required
3~5 mg purified monoclonal antibody needed

Phase I Generation of antibody immunoconjugates
         • Exchange the antibody buffer to the conjugation buffer
         • Activation of the antibody
         • Conjugation of the antibody to TWO different drugs at TWO different molar ratio
         • Purification of the immunoconjugates
         • Filter-sterilization

Phase II In vitro Bioassay
        • Seeding the cells in 96-well plate, up to THREE different cell lines
        • Adding the immunoconjugates at various concentrations.
        • Performing MTT assay on day 5.
        • Repeat the bioassay twice

2. Medium-size Production

Material required
5~10 mg purified monoclonal antibody needed

Phase I Generation of antibody immunoconjugates
        • Exchange the antibody buffer to the conjugation buffer
        • Activation of the antibody
        • Conjugation of the antibody to ONE drug at TWO different molar ratio
        • Purification of the immunoconjugates
        • Filter-sterilization

Phase II In vitro Bioassay
        • Seeding the cells in 96-well plate, up to THREE different cell lines
        • Adding the immunoconjugates at various conc.
        • Performing MTT assay on day 5.
        • Repeat the bioassay once
        • Sending out 2 mg immunoconjugate.

Phase III In vivo animal study
        • Innoculate 30 nude mice with the right number of tumor cells
        • Once the tumors reach 10 mm, 5 mice per group will be injected with various amount of immunoconjugate, naked antibody, unconjugated drug and buffer
        • Tumor size will be measured every 3 or 4 days
        • At the end of the test, the tumors will be removed and weighed individually.
        • Repeat the animal study twice.
 


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