Phage Display, a powerful technique for developing nanobodies.

LOSINNO AD; ÁVILA MANIERO M; DAMIANI MT ; NOLLY MB

Mendoza

Simposio; II International Symposium on Translational Medicine. XIII Annual Meeting of Investigation at the Faculty of Medical Sciences.; 2019

Phage Display, a powerful technique for developing nanobodies.Introduction and objectiveAntibodies are indispensable tools for the recognition of specific proteins termed antigens; used on a daily basis in laboratories, both for diagnosis, analysis, and research. They are currently a cutting-edge therapeutic tool in cancer treatment. Although with different quantitative and qualitative requirements, they are also important in industrial areas. In Argentina, most of the companies that commercialize antibodies do not own their production; therefore they import them, generating an important impact in the overall National Economy.We propose to obtained antibodies by using a novel technology called "Phage Display", based on the variable regions of the immunoglobulin?s known as VHH or nanobodies, which will allow us to obtain antibodies of excellent quality in shorter times and lower costs than the traditional methods.MethodsThe Tomlinson I and J library, a human synthetic VH+VL scFv phage library in a phagemid vector, and the positive control Escherichia coli TG1 (TG1 containing an anti-thyroglobulin clone) were purchased in Source BioScience (Nottingham, England). We followed the protocol provided by the library. Western blot analysis was performed using proteins from dendrites cells, at 10% SDS-PAGE, with anti-Myc antibody followed by HRP conjugated anti-rabbit antibody. Protein detection was performed using an enhanced chemiluminescence assay.Results and Conclusions Two antibodies were obtained, one of them is anti-ubiquitin and the other one is anti-β-galactosidase. Nanobody specificity was confirmed by western blot as described before. "Phage Display" is a highly valued and versatile technology that generates new opportunities for the development and production of monoclonal antibodies without using animals.