title: Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography.
abstract: 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.
authors.affiliation: University of Zagreb, Centre for Research and Knowledge Transfer in Biotechnology, Rockefellerova 10, HR-10000 Zagreb, Croatia; Center of Excellence for Viral Immunology and Vaccines, CERVirVac, Croatia. Electronic address: mbrgles@gmail.com.
authors.forename: Marija
authors.initials: M
authors.lastname: Brgles
authors.affiliation: University of Zagreb, Centre for Research and Knowledge Transfer in Biotechnology, Rockefellerova 10, HR-10000 Zagreb, Croatia; Center of Excellence for Viral Immunology and Vaccines, CERVirVac, Croatia.
authors.forename: Dora
authors.initials: D
authors.lastname: Sviben
authors.affiliation: University of Zagreb, Centre for Research and Knowledge Transfer in Biotechnology, Rockefellerova 10, HR-10000 Zagreb, Croatia; Center of Excellence for Viral Immunology and Vaccines, CERVirVac, Croatia.
authors.forename: Dubravko
authors.initials: D
authors.lastname: Forčić
authors.affiliation: University of Zagreb, Centre for Research and Knowledge Transfer in Biotechnology, Rockefellerova 10, HR-10000 Zagreb, Croatia; Center of Excellence for Viral Immunology and Vaccines, CERVirVac, Croatia.
authors.forename: Beata
authors.initials: B
authors.lastname: Halassy
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity / Q000379:methods; D006863:Hydrogen-Ion Concentration; D009109:Mumps virus / Q000276:immunology / Q000302:isolation & purification*; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification*; D014709:Vero Cells
chemical_list: D000596:Amino Acids; D000906:Antibodies; D011506:Proteins
keywords: Immunoaffinity chromatography; Monolith; Native elution; Virus