Purification of a mouse monoclonal antibody against Erns protein from classical swine fever virus
- Journal of Bacteriology & Mycology: Open Access
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Jose Miguel Fernandez Torres,1 Alianny Lazara Rojo Sanchez,2 Onel Valdivia Perez,1 Reinaldo Blanco Aguila,1 Dayamí Dorta Hernandez,1 Carlos Hernandez,1 Yeosvany Cabrera Artiles,1 Dalia Borges Pérez,1 Daymí Abreu Remedio1
Abstract
With the aim to follow large scale production of recombinant Erns from Classical Swine Fever Virus, a monoclonal specific antibody was obtained and optimized it downstream lab scale purification. The anti-Erns gamma globulins were purified through Protein A and Protein G affinity chromatography from ascites fluid. Three binding factors were assessed (pH, flow rate and buffer) to define dynamic binding capacity. The best binding condition to Protein a (6.6 mg/mL of matrix) was in phosphate buffer, pH 8.0 and linear flow rate of 78cm/h with a purity of 90 %. For Protein G, the best arrangement of factors was phosphate buffer, pH 7.0 and linear flow 178cm/h with 6.45mg/mL of adsorbed antibody with a purity of 95 %. The nProtein-A Sepharose matrix allowed 1.21 times more recovery than the nProtein-G Sepharose matrix. However, the elutions obtained with the G-protein were more pure.
Keywords
swine fever virus, monoclonal antibody, Erns, glycoprotein, biological marker, hybridoma
