Study on ion exchange chromatography condition for purification of Sabin strain polio virus

Abstract

Objective To study on ion exchange chromatography (IEC) condition for purification of Sabin strain polio virus (sPV). Methods The crude purified liquids of sPV obtained by gel chromatography were subjected to IEC in different conditions, and setting parameters included chromatography medium Q Sepharose FF or Eshmuno Q, loading sample volume 30% or 40% column volume (CV), and flow velocity (150±5) or (240±8) cm/h. D antigen contents, protein concenrations, virus titers, host cell protein(HCP)residues and Vero cell DNA residues in purified liquids of sPV purified by IEC were detected, and the recovery rates and specific activities of D antigen were caculated. Results The D antigen recovery rates of the Eshmuno Q IEC purified liquids of sPV were all higher than those of Q Sepharose FF IEC purified. In the conditions of loading sample volume 40% CV and flow velocity (150±5) cm/h, the D antigen specific activities of the Eshmuno Q IEC purified liquids of sPV were all higher than those of Q Sepharose FF IEC purified, and the differences were all statistically significant (typeⅠ: t=4.23, typeⅡ: t=5.73, type Ⅲ: t=4.18, all P values<0.05), and HCP residues (typeⅠ: t=8.29, typeⅡ: t=7.89, type Ⅲ: t=8.18, all P values <0.05) and Vero cell DNA residues (typeⅠ: t=4.23, typeⅡ: t=4.56, type Ⅲ: t=4.78, all P values <0.05) of the former were all lower than those of the latter, all with statistically significant differences. Conclusion When sPV is purified with IEC, replacing Q Sepharose FF with Eshmuno Q can increase loading sample volume and flow velocity of IEC, thus the time of IEC is shortened and the purification efficiency is improved. Key words: Chromatography, ion exchange; Poliovirus vaccine, inactivated; Anion exchange resins