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Pregled bibliografske jedinice broj: 884520

Chromatographic purification of mumps and measles virus


Brgles, Marija
Chromatographic purification of mumps and measles virus // 17th International Chromatography School : Book of abstracts / Ašperger, Danijela ; Bolanča, Tomislav ; Ukić, Šime (ur.).
Zagreb: Fakultet kemijskog inženjerstva i tehnologije Sveučilišta u Zagrebu, 2017. str. 10-11 (pozvano predavanje, domaća recenzija, sažetak, znanstveni)


CROSBI ID: 884520 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Chromatographic purification of mumps and measles virus

Autori
Brgles, Marija

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

Izvornik
17th International Chromatography School : Book of abstracts / Ašperger, Danijela ; Bolanča, Tomislav ; Ukić, Šime - Zagreb : Fakultet kemijskog inženjerstva i tehnologije Sveučilišta u Zagrebu, 2017, 10-11

ISBN
978-953-6470-83-9

Skup
International Chromatography School (17 ; 2017)

Mjesto i datum
Zagreb, Hrvatska, 06.07.2017. - 07.07.2017

Vrsta sudjelovanja
Pozvano predavanje

Vrsta recenzije
Domaća recenzija

Ključne riječi
Chromatography, Virus, Monolith, NanoSight

Sažetak
Mumps and measles virus (MuV and MeV, respectively) are negative strand RNA viruses and belong to the Paramyoviridae family. Both viruses are used for production of live attenuated vaccines used for prophylaxis of mumps and measles disease, respectively. In addition, MeV is a promising gene vector and exhibits oncolytic properties. Production of vaccines and gene therapy biologicals requires virus preparations of highest purity that enable potency and safety of the medicine. Viruses are produced using cell cultures and impurities in crude virus harvests originate either from host cells (e.g. host cell proteins, cellular DNA, exosomes) and cultivation medium (e.g. bovine serum albumin), from processing (e.g. extractables and leachables) or from the virus itself (e.g. virus aggregates, empty capsids, defective interfering particles). Purification of virus particles is still a challenging task due to the complex structure and sensitivity of the viruses. Chromatography is gaining increased interest as a method of virus purification, especially monolith chromatography due to properties beneficial for viruses ; mass transport based on convection, very high porosity, and high binding capacity for very large molecules. Monoliths have already been successfully used for purification and concentration of various viruses, mostly in ion-exchange chromatography. Focus of our research is on MuV and MeV purification and to this aim we have tested immunoaffinity (IAC), ion-exchange (IEX) and hydrophobic interaction chromatography (HIC) on monolith columns. Virus purification efficiency was monitored by measuring concentration of infective particles (CCID50 assay) and total particle concentration (Nanoparticle Tracking Analysis), host cell proteins (measured by ELISA), and genomic host cell DNA (measured by PCR). IAC was found to be very successful with recovery of infective particles around 70 %. Also, we developed a new elution system in IAC under native elution conditions enabling high recoveries of infective virus particles. IAC is very specific and powerful but requires access to specific antibodies, and the use of antibodies requires more quality control which all increase costs of production. Recoveries of infective particles in IEX were quite low, only up to 20 %, and on the other hand recoveries in HIC were around 60 %. Interestingly, total-to- infective particle ratio in the starting sample was found as an important factor in HIC recovery. Channel size of monolith columns was found important in all three investigated types of chromatography. All obtained results are important for biotechnology, but also in fundamental research that requires high purity of virus particles such as study of virus proteome and lipidome.

Izvorni jezik
Engleski

Znanstvena područja
Kemija



POVEZANOST RADA


Projekti:
HRZZ-UIP-2013-11-8193 - Kromatografsko pročišćavanje biomolekula i njihova karakterizacija (CHROBIO) (Brgles, Marija, HRZZ - 2013-11) ( CroRIS)

Ustanove:
Sveučilište u Zagrebu

Profili:

Avatar Url Marija Brgles (autor)


Citiraj ovu publikaciju:

Brgles, Marija
Chromatographic purification of mumps and measles virus // 17th International Chromatography School : Book of abstracts / Ašperger, Danijela ; Bolanča, Tomislav ; Ukić, Šime (ur.).
Zagreb: Fakultet kemijskog inženjerstva i tehnologije Sveučilišta u Zagrebu, 2017. str. 10-11 (pozvano predavanje, domaća recenzija, sažetak, znanstveni)
Brgles, M. (2017) Chromatographic purification of mumps and measles virus. U: Ašperger, D., Bolanča, T. & Ukić, Š. (ur.)17th International Chromatography School : Book of abstracts.
@article{article, author = {Brgles, Marija}, year = {2017}, pages = {10-11}, keywords = {Chromatography, Virus, Monolith, NanoSight}, isbn = {978-953-6470-83-9}, title = {Chromatographic purification of mumps and measles virus}, keyword = {Chromatography, Virus, Monolith, NanoSight}, publisher = {Fakultet kemijskog in\v{z}enjerstva i tehnologije Sveu\v{c}ili\v{s}ta u Zagrebu}, publisherplace = {Zagreb, Hrvatska} }
@article{article, author = {Brgles, Marija}, year = {2017}, pages = {10-11}, keywords = {Chromatography, Virus, Monolith, NanoSight}, isbn = {978-953-6470-83-9}, title = {Chromatographic purification of mumps and measles virus}, keyword = {Chromatography, Virus, Monolith, NanoSight}, publisher = {Fakultet kemijskog in\v{z}enjerstva i tehnologije Sveu\v{c}ili\v{s}ta u Zagrebu}, publisherplace = {Zagreb, Hrvatska} }




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