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Pregled bibliografske jedinice broj: 819415

Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography


Brgles, Marija; Sviben, Dora; Forčić, Dubravko; Halassy, Beata
Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography // 7th Monolith Summer School & Symposium : Book of Abstracts
Portorož, Slovenija, 2016. str. 57-57 (poster, međunarodna recenzija, sažetak, znanstveni)


CROSBI ID: 819415 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography

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

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

Izvornik
7th Monolith Summer School & Symposium : Book of Abstracts / - , 2016, 57-57

Skup
Monolith Summer School & Symposium (7 ; 2016)

Mjesto i datum
Portorož, Slovenija, 29.05.2016. - 01.06.2016

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Immunoaffinity chromatography ; Native elution ; Virus ; Monolith

Sažetak
Immunoaffinity chromatography enables specific purification of any antigen by the use of the antibody raised against it, but also requires quite harsh elution conditions which might affect not only the intermolecular but also the intramolecular interactions. Therefore, the aim of this work was to find conditions that disrupt antigen-antibody interaction without affecting protein structure and stability. We hypothesized that solution of amino acids that are readily found at the antigen-antibody interaction site could act as a competing agent at neutral pH and result in elution of the antigen from the immunoaffinity column. We tested this hypothesis using two immunoaffinity columns ; specific for ovalbumin and mumps virus, and also using protein G affinity column. Immunoaffinity columns were prepared by immobilization of antibodies to epoxy activated monolith columns. Testing of elution potential of amino acids was constrained by limited solubility of amino acids. Results showed that the most efficient elution solutions were those containing imidazole and arginine and their efficiency increases with increasing molarity. Elution efficiency in proteins immunoaffinity chromatography was up to 40 %, but was shown to be beneficial in comparison to standard low pH elution regarding polymer content since amino acid eluates were polymer free in contrast to low pH eluates. Elution efficiency of mumps virus was up to 68 % and separation of noninfective virus particles was achieved. Amino acid solutions at neutral pH show modest elution efficiencies but combined with their stabilizing effect provide new possibilities of immunoaffinity chromatography use in protein and virus purification. *M. Brgles, D. Sviben, D. Forčić, B. Halassy, J. Chromatogr., A (2016) doi: 10.1016/j.chroma.2016.04.022

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 Beata Halassy (autor)

Avatar Url Marija Brgles (autor)

Avatar Url Dubravko Forčić (autor)


Citiraj ovu publikaciju:

Brgles, Marija; Sviben, Dora; Forčić, Dubravko; Halassy, Beata
Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography // 7th Monolith Summer School & Symposium : Book of Abstracts
Portorož, Slovenija, 2016. str. 57-57 (poster, međunarodna recenzija, sažetak, znanstveni)
Brgles, M., Sviben, D., Forčić, D. & Halassy, B. (2016) Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography. U: 7th Monolith Summer School & Symposium : Book of Abstracts.
@article{article, author = {Brgles, Marija and Sviben, Dora and For\v{c}i\'{c}, Dubravko and Halassy, Beata}, year = {2016}, pages = {57-57}, keywords = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, title = {Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography}, keyword = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, publisherplace = {Portoro\v{z}, Slovenija} }
@article{article, author = {Brgles, Marija and Sviben, Dora and For\v{c}i\'{c}, Dubravko and Halassy, Beata}, year = {2016}, pages = {57-57}, keywords = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, title = {Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography}, keyword = {Immunoaffinity chromatography, Native elution, Virus, Monolith}, publisherplace = {Portoro\v{z}, Slovenija} }




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