Cell Based Assays
Antibody Dependent Cellular Cytotoxicity (ADCC), Complement Dependent Cytotoxicity (CDC) & Cell Mediated Cytotoxicity (CMC)
Cell Technology possesses a customizable ADCC/CDC/CMC assay platform that facilitates the selection of antibody drug candidates. Our bioluminescent ultra sensitive aCella-TOX Assay, is rapidly becoming the standard for ADCC/CDC/CMC assays within the therapeutics industry. Please click here for a direct comparison between the aCella-TOX and (51Cr) Chromium Release Methods
We provide ADCC/CDC assay services to determine the activity of the antibody and/or complement against specified target cell lines . The result would be, a dose dependent kinetic response of the therapeutic antibody. The assay service includes optimization of E/T ratios, measurement of ADCC / CDC activity at a series of antibody concentrations, at a certain E:T ratio. Correlation data with FACS analysis can be generated upon request.
Figure: 5000 Ramos cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2 . The ADCC reaction was further incubated for 2 hours (Left graph) or 3.5 hours (Right graph) at the specified E:T ratios. % Cytotoxicity was measured using the aCellaTOX assay. 3.5hour timepoint has been optimized with the 10:1 E:T ratio (Green). Graph with 20:1 E:T ratio (Red) is flat.(100% cytotoxicity)
Figure (Left): 5000 Daudi cells/ well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells maintained overnight with IL-2. The ADCC reaction was further incubated for 2 hours at the specified E:T ratios (25:1 Black and 10:1 Red). % cytotoxicity was measured using the aCellaTOX assay.
Figure (Right): The above graph shows an antibody titration of Rituximab from0.000977 to 1ug/ml incubated with 5% baby rabbit complement and 5000 Daudi target cells for 30 minutes at 37oC
Steps Involved for the ADCC / CDC implementation
Determination of optimal E:T ratios for the Target Cell line.
Procurement of the Antibody (or Antibodies) and Target Cell line (s) to be tested.
A dose response curve would be generated using aCella-TOX or FACS analysis or both. (as requested).
Supplies and information to be provided by the client
Target Cell line(s) (adherent or suspension) (frozen vial(s)) and sufficient Antibody for the project.
Cell Culture Media and conditions for growth of the target cell line
Antibody concentrations (if known)
Number of PBMC donors to be tested.
Information on Antigen expression Level on the target cell line (s) (FACS analysis can be done at the request of the client to determine this).
Supplies to be provided by Cell Technology
All additional supplies would be procured by Cell Technology, and billed to the client at cost.
Final Report would be turned in after the Project with a dose response curve, showing Antibody dependence.
Typical time frame is approximately 4 weeks / Cell line / Antibody.
For Schedule and Pricing, please call 888-727-7297 or 650-960-2170, or email us at [email protected]