2026-05-30T17:49:10Z https://www.peertechzpublications.org/oai-pmh

oai:www.peertechzpublications.org:10.17352/jbm.000007 2019-04-19 PTZ.JBM:VOL3

Different Strategies used in the Production of human monoclonal scfv Antibodies Specific to Dimers of Membrane Receptors Sylwia Łukasiewicz Agata Stachowicz Ewelina Fic Ewa Błasiak Agata KowalikMarta Dziedzicka-Wasylewska<p>Phage display technology is one of the methods widely adopted for the production of human monoclonal antibodies. The subject of the present research was the scFv - single chain variable fragment antibody - which was able to recognize the D2-5-HT1A heteromer formed by dopamine D2 and serotonin 5-HT1A receptors. The antibody was selected using the phage display technique and the human antibody scFv phagemid library Tomlinson J. We investigated two expression systems: prokaryotic and eukaryotic. The results indicate that both of these systems (E.coli HB2151 and High Five cells (BTI-TN-5B1-4) of Trichopulsia ni) are suitable for the production of scFv antibodies, but the scFvs obtained from insect cells could attain a higher protein concentration. Next, we examined a few purification methods: immobilized-metal affinity chromatography (IMAC), affinity chromatography with the c-myc tag and the methods using protein L or A agarose, which are applicable to scFvs lacking tags. In the bacterial as well as insect cell expression systems, protein L agarose chromatography gave satisfactory results, and in our opinion, it is the best choice as a purification resin. Additionally, circular dichroism measurements confirmed the predominant content of secondary structures in the tested antibodies, which is consistent with the literature data.</p> Journal of Biology and Medicine - Peertechz Publications 2019-04-19 Research Article https://doi.org/10.17352/jbm.000007 en Copyright © Sylwia Łukasiewicz et al.