LARGE-SCALE PRODUCTION AND CHARACTERIZATION OF THE EXOSOME TRANSPORTER SYSTEM FROM THP-1 CELLS AS A NEW APPROACH TO IMMUNIZATION
Bildiriler > Özet Bildiri · 2022
Yazarlar: Ilgın Kımız Geboloğlu, Süleyman Furkan Demirden, Ayşe Gülten Kantarcı, Suphi Şurişvan ÖNCEL
4th International Vaccinology Congress, Türkiye, Ankara, Uluslararası, Sözlü Sunum, Özet Bildiri, 2022-2022, cilt ., ss 61-61
DOI: .
Özet
<p>The development of effective vaccines against many life-threatening diseases is one of the most important medical achievements in recent history. In immunization created by vaccination, it is crucial that the appropriate antigens can be processed efficiently by the immune system. Therefore, in recent years, there has been a trend toward lipid-based nanoparticles targeting the mimicry of cellular mechanisms in the body. The use of exosomes, called natural liposomes, which are membrane vesicles released from prokaryotic and eukaryotic cells, such as microvesicles and apoptotic bodies, as carrier systems is becoming an increasingly exciting area. The aim of this study is large-scale production in a controlled bioreactor, isolation, and characterization of exosomes which can be used as carrier systems in immunization. In this way, great quantities of exosomes can be obtained in a short time. Therefore, this can be beneficial for their commercial applicability by considering their potential usage areas in biopharmaceutical industries. Since the target usage area for these exosomes is biopharmaceutical, THP-1 (human monocyte) cells were adapted to a chemically defined medium (FBCD, Florabio) that does not contain any animal-derived compounds. Thus, unwanted animal originated exosomes are also prevented. Then, serum-free medium adapted cells were optimized in erlenmayers and spinner flasks before production in a stirred tank bioreactor (STR). The ultrafiltration method, which is more suitable for large-scale production, was used for the isolation of exosomes. After, isolated exosomes were characterized in size, homogeneity, and protein concentration. THP-1 cells were gradually adapted to the FBCD medium. It was determined that the initial cell concentration should be 5x105 cells/mL, the stirring speed should be 70 rpm in the spinner with a working volume of 100 mL and 1.85x106 cells/mL was obtained under these conditions. As a result of exosome isolation from the spinner flask, exosomes were obtained with a size of 74.95 ± 2.15 nm and an exosomal protein concentration of 360.00 ± 8.08 μg/mL. In STR production, 2.06x106 cells/mL concentration cells were obtained and after isolation, exosomes were obtained with a size of 71.52 ± 1.98 nm and an exosomal protein concentration of 468.57 ± 2.02 μg/mL. Detailed characterization studies of the exosomes are ongoing.</p><p>Acknowledgments: This work was supported through research grants from Ege University Scientific Research Projects Coordination with a project number of 22868.</p><p>Keywords: exosomes, stirred tank bioreactor, lipid-based delivery system, vaccination, immunization</p>