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Pregled bibliografske jedinice broj: 833303

Stability of proteins and viruses determined by differential scanning fluorimetry


Sviben, Dora; Forčić, Dubravko; Halassy, Beata; Zurawski, Marek; Brgles, Marija
Stability of proteins and viruses determined by differential scanning fluorimetry // 13th International "Greta Pifat Mrzljak" School of Biophysics "ABC of Physics of Life" : Book of abstracts / Marion, Sanjin ; Delač Marion, Ida ; Maltar Strmečki, Nadica ; Josef-Golubić, Sanja ; Vuletić, Tomislav (ur.).
Zagreb: Croatian Biophysical Societey & Institute of Physics, 2016. str. 145-145 (predavanje, međunarodna recenzija, sažetak, znanstveni)


CROSBI ID: 833303 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Stability of proteins and viruses determined by differential scanning fluorimetry

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

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

Izvornik
13th International "Greta Pifat Mrzljak" School of Biophysics "ABC of Physics of Life" : Book of abstracts / Marion, Sanjin ; Delač Marion, Ida ; Maltar Strmečki, Nadica ; Josef-Golubić, Sanja ; Vuletić, Tomislav - Zagreb : Croatian Biophysical Societey & Institute of Physics, 2016, 145-145

ISBN
978-953-7666-14-9

Skup
International "Greta Pifat Mrzljak" School of Biophysics "ABC of Physics of Life" (13 ; 2016)

Mjesto i datum
Hrvatska, 01.09.2016. - 10.09.2016

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
protein ; virus ; differential scanning fluorimetry ; SEC-HPLC

Sažetak
Differential scanning fluorimetry (DSF) is readily employed high-throughput method for probing protein stability. Typically, DSF monitors thermal unfolding of proteins using a fluorescent reporter dye (e.g. Sypro Orange) and a real-time PCR instrument[1]. Recently, a new method, nanoDSF, was developed which enables measuring of intrinsic tryptophan and tyrosine fluorescence[2, 3], its main advantage being that no fluorescent dye is needed, thus avoiding any detrimental dye-protein interactions. Both in DSF and nanoDSF, the fluorescence intensity increases with the exposure of hydrophobic protein parts. When plotted as a function of temperature, it gives a value of Tm at which the concentrations of folded and unfolded proteins are equal. Effect of different compounds on protein stability is seen as a change in Tm. Stability of human immunoglobulin G (hIgG) and human serum albumin (HSA) is very interesting because of their therapeutical use. DSF and nanoDSF were used to examine the stability of hIgG, HSA and ovalbumin (OVA) in different formulations (e.g. pH, salts, amino acids, sugars) and the methods showed excellent correlation. Formulations containing ammonium sulphate and sucrose proved as best stabilizers, while pronounced destabilizing effects were observed with imidazole, arginine and at low pH (pH 4). Thermal stability of measles and mumps virus was also probed, but several problems occurred – acquiring freshly concentrated viruses, damage to the viruses during concentration and lack of clear interpretation of the obtained Tm. Protein stability was also monitored by measuring protein aggregation by SEC-HPLC and virus stability by CCID50. Obtained results correlate well with DSF results. [1] Niesen FH, Berglund H, Vedadi M (2007) The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability. Nat Protoc 2:2212–2221. [2] http://www.nanotemper- technologies.com/technologies/nanodsf/ (accessed on 21-06-2016). [3] Temel DB, Landsman P, Brader ML (2016) Orthogonal methods for characterizing the unfolding of therapeutic monoclonal antibodies: differential scanning calorimetry, isothermal chemical denaturation, and intrinsic fluorescence with concomitant static light scattering. Methods Enzymol 567:359–389.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

Napomena
Rad je na istom skupu prezentiran i kao poster.



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:

Sviben, Dora; Forčić, Dubravko; Halassy, Beata; Zurawski, Marek; Brgles, Marija
Stability of proteins and viruses determined by differential scanning fluorimetry // 13th International "Greta Pifat Mrzljak" School of Biophysics "ABC of Physics of Life" : Book of abstracts / Marion, Sanjin ; Delač Marion, Ida ; Maltar Strmečki, Nadica ; Josef-Golubić, Sanja ; Vuletić, Tomislav (ur.).
Zagreb: Croatian Biophysical Societey & Institute of Physics, 2016. str. 145-145 (predavanje, međunarodna recenzija, sažetak, znanstveni)
Sviben, D., Forčić, D., Halassy, B., Zurawski, M. & Brgles, M. (2016) Stability of proteins and viruses determined by differential scanning fluorimetry. U: Marion, S., Delač Marion, I., Maltar Strmečki, N., Josef-Golubić, S. & Vuletić, T. (ur.)13th International "Greta Pifat Mrzljak" School of Biophysics "ABC of Physics of Life" : Book of abstracts.
@article{article, author = {Sviben, Dora and For\v{c}i\'{c}, Dubravko and Halassy, Beata and Zurawski, Marek and Brgles, Marija}, year = {2016}, pages = {145-145}, keywords = {protein, virus, differential scanning fluorimetry, SEC-HPLC}, isbn = {978-953-7666-14-9}, title = {Stability of proteins and viruses determined by differential scanning fluorimetry}, keyword = {protein, virus, differential scanning fluorimetry, SEC-HPLC}, publisher = {Croatian Biophysical Societey and Institute of Physics}, publisherplace = {Hrvatska} }
@article{article, author = {Sviben, Dora and For\v{c}i\'{c}, Dubravko and Halassy, Beata and Zurawski, Marek and Brgles, Marija}, year = {2016}, pages = {145-145}, keywords = {protein, virus, differential scanning fluorimetry, SEC-HPLC}, isbn = {978-953-7666-14-9}, title = {Stability of proteins and viruses determined by differential scanning fluorimetry}, keyword = {protein, virus, differential scanning fluorimetry, SEC-HPLC}, publisher = {Croatian Biophysical Societey and Institute of Physics}, publisherplace = {Hrvatska} }




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