Pregled bibliografske jedinice broj: 808750
Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography
Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography // Journal of chromatography. A, 1447 (2016), 107-114 doi:10.1016/j.chroma.2016.04.022 (međunarodna recenzija, članak, znanstveni)
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Naslov
Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography
Autori
Brgles, Marija ; Sviben, Dora ; Forčić, Dubravko ; Halassy, Beata
Izvornik
Journal of chromatography. A (0021-9673) 1447
(2016);
107-114
Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni
Ključne riječi
Immunoaffinity chromatography ; Native elution ; Virus ; Monolith
Sažetak
Immunoaffinity chromatography, based on the antigen-antibody recognition, enables specific purification of any antigen (protein, virus) by its antibody. The problem with immunoaffinity chromatography is the harsh elution conditions required for disrupting strong antigen-antibody interactions, such as low pH buffers, which are often deleterious for the immobilized protein and the protein to be isolated since they can also disrupt the intramolecular forces. Therefore, immunoaffinity chromatography can only be partially used for protein and virus purification. Here we report on a nonspecific elution in immunoaffinity chromatography using native conditions by elution with amino acid solution at physiological pH for which we suppose possible competing mechanism of action. Elution potential of various amino acid solutions was tested using immunoaffinity columns specific for ovalbumin and mumps virus, and protein G affinity column. Results have shown that the most successful elution solutions were those containing imidazole and arginine of high molarity. Imidazole represents aromatic residues readily found at the antigen- antibody interaction surface and arginine is most frequently found on protein surface in general. Therefore, results on their eluting power in immunoaffinity chromatography, which increases with increasing molarity, are in line with the competing mechanism of action. Virus immunoaffinity chromatography resulted in removal on nonviable virus particles, which is important for research and biotechnology purposes. In addition, amino acids are proven stabilizers for proteins and viruses making approach presented in this work a very convenient purification method.
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
Citiraj ovu publikaciju:
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