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Selection and Characterization of Single Chain Antibody Fragments Specific for Hsp90 as a Potential Cancer Targeting
Edyta Petters1, Aleksandra Sokolowska-Wedzina2, Jacek Otlewski3,4
1Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. edyta_petters@op.pl.
Abstract:
Heat shock proteins play an essential role in facilitating malignant transformation and they have been recognized as important factors in human cancers. One of the key elements of the molecular chaperones machinery is Hsp90 and it has recently become a target for anticancer therapeutic approaches. The potential and importance of Hsp90-directed agents becomes apparent when one realizes that disruption of Hsp90 function may influence over 200 oncogenic client proteins. Here, we described the selection and characterization of Hsp90-specific antibody fragments from commercially available Tomlinson I and J phage display libraries. The affinities of Hsp90-binding scFv variants were measured using SPR method. Then, based on the best clone selected, we performed the affinity maturation procedure and obtained valuable Hsp90-specific clones. The selected binders were expressed and applied for immunostaining, ELISA and SPR analysis using model cancer cell lines. All performed experiments confirmed the ability of selected antibodies to interact with the Hsp90. Therefore, the presented Hsp90-specific scFv, might be a starting point for the development of a novel antibody-based strategy targeting cancer.
Insights
Researchers developed novel Hsp90-specific antibody fragments for cancer therapy. These single-chain variable fragments (scFv) show potential for targeting heat shock protein 90 (Hsp90) in cancer treatment strategies.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Heat shock proteins (HSPs), particularly Hsp90, are crucial for cancer cell survival and proliferation.
- Hsp90 is a validated target for anticancer therapies due to its role in stabilizing over 200 oncogenic client proteins.
Purpose of the Study:
- To select and characterize novel Hsp90-specific antibody fragments.
- To develop potential tools for antibody-based cancer therapeutics targeting Hsp90.
Main Methods:
- Utilized phage display libraries (Tomlinson I and J) for antibody fragment selection.
- Employed Surface Plasmon Resonance (SPR) for affinity measurement and characterization.
- Performed affinity maturation to enhance antibody binding.
- Expressed and validated antibody fragments using immunostaining, ELISA, and SPR on cancer cell lines.
Main Results:
- Successfully selected and characterized Hsp90-specific single-chain variable fragment (scFv) antibodies.
- Demonstrated high affinity and specific binding of selected scFv to Hsp90.
- Confirmed the interaction of antibodies with Hsp90 in model cancer cell lines.
Conclusions:
- The developed Hsp90-specific scFv antibodies represent promising candidates for further development.
- These antibody fragments could serve as a foundation for novel antibody-based cancer treatment strategies targeting Hsp90.
