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Cloning and expression of a novel murine anti-human Fas antibody
H Yoshida-Kato1, K Ichikawa, J Yamaguchi
1Biomedical Research Laboratories, Sankyo Co, Ltd, Tokyo, Japan.
Abstract:
Agonistic anti-human Fas antibodies that can induce apoptosis are thought to have therapeutic effects for various diseases resulting from an abnormality of the Fas/FasL system. However, some anti-Fas antibodies show toxicity, and it is difficult to investigate their therapeutic and toxicological effect using animals because of their species specificity. We previously obtained a murine anti-human Fas mAb, HFE7A. HFE7A reacted with both human and murine Fas, and mitigated lymphadenopathy without any sign of hepatotoxicity in MRLgld/gld mice. It is suggested that humanized HFE7A would be a therapeutic treatment for various diseases resulting from an abnormality of the Fas/FasL system. Here we isolated the cDNAs that code for the heavy and light chains of HFE7A and identified the corresponding nucleotide sequences. The recombinant HFE7A was indistinguishable in binding and apoptosis-inducing activity to that from a hybridoma cell line. These data provide essential information for the humanization and clinical application of the humanized HFE7A.
Insights
Researchers developed a novel murine anti-human Fas antibody (HFE7A) that targets Fas/FasL system abnormalities. This antibody mitigated disease in mice without toxicity, paving the way for humanized therapeutic applications.
Area of Science:
- Immunology
- Molecular Biology
- Therapeutics
Background:
- Agonistic anti-Fas antibodies show therapeutic potential for Fas/FasL system disorders.
- Species specificity of anti-Fas antibodies complicates therapeutic and toxicological studies in animal models.
- Murine anti-human Fas mAb HFE7A demonstrated efficacy and safety in preclinical models.
Purpose of the Study:
- To isolate and characterize the cDNAs encoding the heavy and light chains of the HFE7A antibody.
- To confirm the binding and apoptosis-inducing activity of recombinant HFE7A.
- To provide data for the humanization and clinical application of HFE7A.
Main Methods:
- Isolation of cDNAs for HFE7A heavy and light chains.
- Nucleotide sequencing of the identified cDNAs.
- Comparison of recombinant HFE7A with hybridoma-derived HFE7A for binding and apoptosis-inducing activity.
Main Results:
- Nucleotide sequences for HFE7A heavy and light chains were identified.
- Recombinant HFE7A exhibited comparable binding and apoptosis-inducing activity to the original antibody.
- The characterized HFE7A provides a foundation for developing humanized therapeutic antibodies.
Conclusions:
- The characterization of HFE7A provides essential data for its humanization.
- Humanized HFE7A holds promise as a therapeutic agent for Fas/FasL-related diseases.
- This research facilitates the clinical application of novel anti-Fas antibody therapies.