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Pregled bibliografske jedinice broj: 851458

Native elution in immunoaffinity chromatography of viruses – a step toward high-purity virus particle purification


Brgles, Marija; Sviben, Dora; Forčić, Dubravko; Halassy, Beata
Native elution in immunoaffinity chromatography of viruses – a step toward high-purity virus particle purification // 2016 Annual Meeting of the Croatian Immunological Society : Abstract Book / Kelava, Tomislav ; Markotić, Antonio ; Šućur, Alan (ur.).
Zagreb: Hrvatsko imunološko društvo, 2016. str. 38-38 (predavanje, domaća recenzija, sažetak, znanstveni)


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Naslov
Native elution in immunoaffinity chromatography of viruses – a step toward high-purity virus particle purification

Autori
Brgles, Marija ; Sviben, Dora ; Forčić, Dubravko ; Halassy, Beata

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

Izvornik
2016 Annual Meeting of the Croatian Immunological Society : Abstract Book / Kelava, Tomislav ; Markotić, Antonio ; Šućur, Alan - Zagreb : Hrvatsko imunološko društvo, 2016, 38-38

Skup
Annual Meeting of the Croatian Immunological Society

Mjesto i datum
Ogulin, Hrvatska, 2016

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Domaća recenzija

Ključne riječi
Immunoaffinity chromatography ; Native elution ; Virus ; Monolith

Sažetak
Whole viral particles are the active principles in prophylactic (attenuated viruses) and therapeutic vaccines (viral vectors). The manufacturing process of viral particles consists of upstream (USP, virus in vivo production in bioreactors) and downstream (DSP, virus purification) part. The immunogenicity and stability of viruses throughout the entire production chain should be maintained. The main aim of DSP is to eliminate contaminants, either process related or product related (host cell proteins, DNA, free proteins, aggregated and empty capsids, host cell exosomes). Downstream processing of viruses has currently been a bottleneck of virus manufacturing, and accounts for up to 70% of overall production costs. Immunoaffinity chromatography is one of the most powerful techniques in the protein purification. High specificity and high affinity of antigen-antibody interaction enables high purification (up to 1000x) and concentration in a single step. However, the drawback of immunoaffinity is that the high affinity interaction between antigen and antibody can be disrupted only under harsh elution conditions that inevitably disturb also intramolecular non-covalent interaction, disordering the protein conformation, and consequently function. Particularly viruses are especially sensitive to elution condition effective in immunoaffinity chromatography, so the immunoaffinity chromatography has not been used in purification of viable viruses. Here we describe the novel principle of native elution, i.e. effective elution of viruses under native, physiological conditions that maintains virus stability and infectivity. It is based on elution of antigen in immunoaffinity chromatography by different amino acid solutions of high molarity, under physiological pH (7.2-7.4). The mechanism of antigen (virus) desorption from antibody is competition. Using attenuated mumps virus strain, as a model virus, we were able to elute 49±16.5% (n=10) infective virus particles using 0.75 M Arg/0.75 M imidazole as eluting agent and 68±13.5% (n=12) infective virus particles using 0.75 M Arg/0.75 M Ser. Moreover, we were able to demonstrate that native elution was effective in separation of infective from non- infective particles. Since amino acids have already been experimentally demonstrated to have virus stabilizing features, a combination of high specificity exhibited by immunoaffinity chromatography and efficient elution with pH neutral, stabilizing solution opens new possibilities for commercial use.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Biotehnologija



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 Beata Halassy (autor)

Avatar Url Marija Brgles (autor)


Citiraj ovu publikaciju:

Brgles, Marija; Sviben, Dora; Forčić, Dubravko; Halassy, Beata
Native elution in immunoaffinity chromatography of viruses – a step toward high-purity virus particle purification // 2016 Annual Meeting of the Croatian Immunological Society : Abstract Book / Kelava, Tomislav ; Markotić, Antonio ; Šućur, Alan (ur.).
Zagreb: Hrvatsko imunološko društvo, 2016. str. 38-38 (predavanje, domaća recenzija, sažetak, znanstveni)
Brgles, M., Sviben, D., Forčić, D. & Halassy, B. (2016) Native elution in immunoaffinity chromatography of viruses – a step toward high-purity virus particle purification. U: Kelava, T., Markotić, A. & Šućur, A. (ur.)2016 Annual Meeting of the Croatian Immunological Society : Abstract Book.
@article{article, author = {Brgles, Marija and Sviben, Dora and For\v{c}i\'{c}, Dubravko and Halassy, Beata}, year = {2016}, pages = {38-38}, keywords = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, title = {Native elution in immunoaffinity chromatography of viruses – a step toward high-purity virus particle purification}, keyword = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, publisher = {Hrvatsko imunolo\v{s}ko dru\v{s}tvo}, publisherplace = {Ogulin, Hrvatska} }
@article{article, author = {Brgles, Marija and Sviben, Dora and For\v{c}i\'{c}, Dubravko and Halassy, Beata}, year = {2016}, pages = {38-38}, keywords = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, title = {Native elution in immunoaffinity chromatography of viruses – a step toward high-purity virus particle purification}, keyword = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, publisher = {Hrvatsko imunolo\v{s}ko dru\v{s}tvo}, publisherplace = {Ogulin, Hrvatska} }




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