Related Experiment Videos
Characterization of human tissue factor (TF)-specific monoclonal antibodies prepared using a rapid immunization
Chaoyuan Chen1, Qifeng Yang, Sima Patel
1Department of Immunotherapeutics, Purdue Pharma L.P., Cranbury, New Jersey, USA.
Hybridoma (2005)
|April 29, 2005
Summary
Researchers developed novel mouse monoclonal antibodies (MAbs) targeting human tissue factor (TF) without affecting blood coagulation. This rapid MAb generation method, using repetitive immunizations at multiple sites (RIMMS), offers a reliable approach for developing targeted therapies.
Area of Science:
- Immunology
- Oncology
- Hematology
Background:
- Tissue factor (TF) is crucial for hemostasis.
- Over-expressed TF is linked to solid tumors, angiogenesis, and metastasis.
- Targeting TF offers potential therapeutic strategies.
Purpose of the Study:
- To develop and characterize murine monoclonal antibodies (MAbs) specific for human TF.
- To generate MAbs that do not inhibit TF-mediated blood coagulation.
- To evaluate the repetitive immunizations at multiple sites (RIMMS) protocol for rapid MAb generation.
Main Methods:
- Utilized a modified repetitive immunizations at multiple sites (RIMMS) protocol.
- Employed hybridoma technology for MAb generation.
- Screened MAbs using ELISA, flow cytometry, and BIAcore analysis.
Main Results:
- Generated a panel of murine MAbs specific for human TF.
- Several MAbs demonstrated high affinity binding (0.5-2 nM) to native human TF.
- Selected MAbs did not inhibit TF-mediated blood coagulation.
- Confirmed the efficiency and reliability of the RIMMS protocol for MAb generation.
Conclusions:
- The RIMMS protocol enables rapid and reliable generation of high-affinity, class-switched MAbs.
- Developed TF-specific MAbs that do not impede blood coagulation, suitable for further therapeutic development.
- This study validates RIMMS as an effective strategy for producing targeted therapeutic antibodies.