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Pregled bibliografske jedinice broj: 950766

Investigation of the thermal shift assay and its power to predict protein and virus stabilizing conditions


Sviben, Dora; Bertoša, Branimir; Hloušek- Kasun, Andrea; Forčić, Dubravko; Halassy, Beata; Brgles, Marija
Investigation of the thermal shift assay and its power to predict protein and virus stabilizing conditions // Journal of pharmaceutical and biomedical analysis, 161 (2018), 73-82 doi:10.1016/j.jpba.2018.08.017 (međunarodna recenzija, članak, znanstveni)


CROSBI ID: 950766 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Investigation of the thermal shift assay and its power to predict protein and virus stabilizing conditions

Autori
Sviben, Dora ; Bertoša, Branimir ; Hloušek- Kasun, Andrea ; Forčić, Dubravko ; Halassy, Beata ; Brgles, Marija

Izvornik
Journal of pharmaceutical and biomedical analysis (0731-7085) 161 (2018); 73-82

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Thermal shift assay ; Excipients ; Biopharmaceutics ; Virus stability ; Protein aggregation

Sažetak
Protein thermal shift assay (TSA) has been extensively used in investigation of protein stabilization (for protein biopharmaceutics stabilization, protein crystallization studies or screening of recombinant proteins) and drug discovery (screening of ligands or inhibitors). This work aimed to analyze thermal shift assay results in comparison to protein polymerization (multimerization and aggregation) propensity and test the most stabilizing formulations for their stabilization effect on enveloped viruses. Influence of protein concentration, buffer pH and molarity was tested on three proteins (immunoglobulin G, ovalbumin, and albumin) and results showed that each of these factors has an impact on determined shift in protein melting point Tm, and the impact was similar for all three proteins. In case of ovalbumin, molecular dynamics simulations were performed with the goal to understanding molecular basis of protein’s thermal stability dependence on pH. Effect of three denaturing agents in a wide concentration range on Tm showed nicely that chemical denaturation occurs only at the highest concentrations. Results showed similar effect on Tm for most formulations on different proteins. Most successful formulations were tested for enveloped virus stabilizing potential using cell culture infectivity assay (CCID50) and results showed lack of correlation with TSA results. Only weak correlation of Tm shift and protein polymerization measured by SEC-HPLC was obtained, meaning that polymerization cannot be predicted from Tm shifts.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biotehnologija



POVEZANOST RADA


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

Ustanove:
Prirodoslovno-matematički fakultet, Zagreb,
Sveučilište u Zagrebu

Citiraj ovu publikaciju

Sviben, Dora; Bertoša, Branimir; Hloušek- Kasun, Andrea; Forčić, Dubravko; Halassy, Beata; Brgles, Marija
Investigation of the thermal shift assay and its power to predict protein and virus stabilizing conditions // Journal of pharmaceutical and biomedical analysis, 161 (2018), 73-82 doi:10.1016/j.jpba.2018.08.017 (međunarodna recenzija, članak, znanstveni)
Sviben, D., Bertoša, B., Hloušek- Kasun, A., Forčić, D., Halassy, B. & Brgles, M. (2018) Investigation of the thermal shift assay and its power to predict protein and virus stabilizing conditions. Journal of pharmaceutical and biomedical analysis, 161, 73-82 doi:10.1016/j.jpba.2018.08.017.
@article{article, year = {2018}, pages = {73-82}, DOI = {10.1016/j.jpba.2018.08.017}, keywords = {Thermal shift assay, Excipients, Biopharmaceutics, Virus stability, Protein aggregation}, journal = {Journal of pharmaceutical and biomedical analysis}, doi = {10.1016/j.jpba.2018.08.017}, volume = {161}, issn = {0731-7085}, title = {Investigation of the thermal shift assay and its power to predict protein and virus stabilizing conditions}, keyword = {Thermal shift assay, Excipients, Biopharmaceutics, Virus stability, Protein aggregation} }
@article{article, year = {2018}, pages = {73-82}, DOI = {10.1016/j.jpba.2018.08.017}, keywords = {Thermal shift assay, Excipients, Biopharmaceutics, Virus stability, Protein aggregation}, journal = {Journal of pharmaceutical and biomedical analysis}, doi = {10.1016/j.jpba.2018.08.017}, volume = {161}, issn = {0731-7085}, title = {Investigation of the thermal shift assay and its power to predict protein and virus stabilizing conditions}, keyword = {Thermal shift assay, Excipients, Biopharmaceutics, Virus stability, Protein aggregation} }

Časopis indeksira:


  • Current Contents Connect (CCC)
  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus
  • MEDLINE


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