Mono- and polyprotic buffer systems in anion exchange chromatography of influenza virus particles

Abstract

Different ions typically used in downstream processing of biologicals are evaluated for their potential in anion exchange chromatography of an industrially produced, pandemic influenza H1N1 virus. Capacity, selectivity and recovery are investigated based on single step elution parallel chromatography experiments. The inactivated H1N1 feedstream is produced in Madin-Darby Bovine Kidney cells. Interesting effects are found for sodium phosphate and sodium citrate. Both anions are triprotic kosmotropes. Anion exchange chromatography generally offers high scalability to satisfy sudden demands for vaccines, which may occur in case of an emerging influenza outbreak. Appropriate pH conditions for H1N1 adsorption are determined by Zeta potential measurements. The dynamic binding capacity of a salt tolerant polyamine-type resin is up to 6.4 times greater than the capacity of a grafted Q-type resin. Pseudo-affinity interactions of polyamines with the M2 protein of influenza may contribute to the obtained capacity increase. Both resins achieve greater capacity in sodium phosphate buffer compared to Tris/HCl. A recovery of 67% and DNA clearance close to 100% without DNAse treatment are achieved for the Q-type resin. Recovery of the virus from the salt tolerant resin requires the use of polyprotic acids in the elution buffer. 85% of the DNA and 60% of the proteins can be removed by the salt tolerant resin. The presence of sodium phosphate during anion exchange chromatography seems to support stability of the H1N1 particles in presence of hydrophobic cations.

Purification of oncolytic measles virus by cation-exchange chromatography using resin-based stationary phases

“…Sodium phosphate buffer instead of Tris/HCl was previously shown to increase the recovery of influenza A virus by 30% during AEX chromatography . [35] However, we did not observe such a profound improvement. The best recovery was again achieved by the S1063 resin, but improved only marginally to 10.3% (Table 2).…”

“…At high flow rates, the size of MV particles and their resistance to diffusion should prevent interactions with the active surfaces of resin pores . [9] However, resin-based stationary phases have achieved the successful purification of influenza A virus, [34,35] nervous necrosis virus [36] and porcine reproductive and respiratory syndrome virus, [37] leading us to believe they may also be suitable for oncolytic MV. Accordingly, we evaluated resin-based IEC with a range of commercial and noncommercial resins in bind-and-elute mode for the purification of oncolytic MV.…”

Safe and effective disinfection of show cave infrastructure in a time of COVID-19

“…1). As influenza includes other surface proteins along with a negatively charged phospholipid bilayer, this obtained value was compared to the measured pI for the entire viral particle, which is 5.0 to 5.9 (Miller et al, 1944;Vajda et al, 2016). Similar to coronaviruses, rabies has a dominant glycoprotein with a calculated pI of 7.6 ( Fig.…”

Towards integrated production of an influenza A vaccine candidate with MDCK suspension cells

“…Techniques for downstream processing (DSP) of virus particles at an industrial scale typically involve filtration and chromatography methods (Wolf & Reichl, 2011). Examples of the latter are ion‐exchange chromatography (IEX) (Lee et al, 2015; Vajda et al, 2016), size exclusion chromatography (SEC) (Kröber et al, 2013; Yuan et al, 2015), hydrophobic interaction chromatography (Li et al, 2015; Wolff et al, 2010), affinity and pseudo‐affinity chromatography (B. Carvalho et al, 2018; Fortuna et al, 2018), and multimodal chromatography (Baek et al, 2011; Kuiper et al, 2002). Standard unit operations, like depth filtration, (ultra‐)centrifugation, (ultra‐)filtration, and column chromatography are generally combined to build a DSP train (Morenweiser, 2005; Wolf & Reichl, 2011; Wolff & Reichl, 2008).…”