title: Selection and Characterization of Single Chain Antibody Fragments Specific for Hsp90 as a Potential Cancer Targeting Molecule.
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.
authors: Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. edyta_petters@op.pl.
authors: Edyta
authors: E
authors: Petters
authors: Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. aleksandra.sokolowska-wedzina@uwr.edu.pl.
authors: Aleksandra
authors: A
authors: Sokolowska-Wedzina
authors: Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. jacek.otlewski@uwr.edu.pl.|Wroclaw Research Centre EIT+, Stablowicka 147, 54-066 Wroclaw, Poland. jacek.otlewski@uwr.edu.pl.
authors: Jacek
authors: J
authors: Otlewski
mesh_terms: D000915:Antibody Affinity; D001943:Breast Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D045744:Cell Line, Tumor; D005260:Female; D018841:HSP90 Heat-Shock Proteins / Q000276:immunology*; D006801:Humans; D058990:Molecular Targeted Therapy; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D018841:HSP90 Heat-Shock Proteins; D019151:Peptide Library; D057127:Single-Chain Antibodies
keywords: Heat Shock Protein 90; antibody fragments; cancer marker; phage display