Naslov
Comparison of production- and purification-relevant properties of human and murine cytomegalovirus

Autori
Ravlić, Sanda ; Brgles, Marija ; Hiršl, Lea ; Jonjić, Stipan ; Halassy, Beata ;

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
Annual Meeting of the Croatian Immunological Society 2021

Mjesto i datum
Trogir, Hrvatska, 23.09.2021. - 25.09.2021

Vrsta sudjelovanja
Poster

Vrsta recenzije
Podatak o recenziji nije dostupan

Ključne riječi
citomagalovirus, HCMV, MCMV, kromatografija ionske izmjene, virus, pročišćavanje
(Cytomegalovirus, HCMV, MCMV, ion exchange chromatography, virus, purification)

Sažetak
The impact of human CMV (HCMV) infections on public health is significant while affecting the most vulnerable groups, immunocompromised individuals and congenitally infected infants. Thus, a vaccine to reduce the incidence and severity of HCMV infection is a public health priority. Moreover, cytomegalovirus has a number of features that makes it a very interesting vector platform for gene therapy. In both cases, preparation of highly purified virus is a prerequisite for safe and effective application. Murine CMV (MCMV) is by far the best studied model for HCMV infections with regard to the principles that govern the immune surveillance of CMVs. The transfer of knowledge from MCMV and mice to HCMV and humans could face challenges not only because of differences in the immune systems of these two species, but also because of differences in the biological and biophysical properties of the two viruses. We carried out a detailed investigation of the MCMV and HCMV growth kinetics as well as stability under the influence of clarification, different storage conditions and ultracentrifugation. We also investigated possibilities to purify both viruses by ion-exchange chromatography. The effectiveness of the procedures was monitored using CCID50 assay, Nanoparticle tracking analysis (NTA), ELISA for host cell proteins, and quantitative PCR assay for host cell DNA. MCMV generally proved to be more robust in handling and despite its greater sensitivity, HCMV was efficiently (100% recovery) purified and concentrated by anion-exchange chromatography using QA monolithic support. These results provide important data for research on all upstream and downstream processes on these two viruses regarding biotechnological production and basic research.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)

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