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Int J Mol Sci. 2015 Aug 21;16(8):19920-35. doi: 10.3390/ijms160819920.

Selection and Characterization of Single Chain Antibody Fragments Specific for Hsp90 as a Potential Cancer Targeting Molecule.

Author information

1
Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. edyta_petters@op.pl.
2
Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. aleksandra.sokolowska-wedzina@uwr.edu.pl.
3
Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. jacek.otlewski@uwr.edu.pl.
4
Wroclaw Research Centre EIT+, Stablowicka 147, 54-066 Wroclaw, Poland. jacek.otlewski@uwr.edu.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.

KEYWORDS:

Heat Shock Protein 90; antibody fragments; cancer marker; phage display

PMID:
26307975
PMCID:
PMC4581332
DOI:
10.3390/ijms160819920
[Indexed for MEDLINE]
Free PMC Article

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