Comparison of Influenza Virus Particle Purification Using Magnetic Sulfated Cellulose Particles with an Established Centrifugation Method for Analytics

Abstract

A method for the purification of influenza virus particles using novel magnetic sulfated cellulose particles is presented and compared to an established centrifugation method for analytics. Therefore, purified influenza A virus particles from adherent and suspension MDCK host cell lines were characterized on the protein level with mass spectrometry to compare the viral and residual host cell proteins. Both methods allowed one to identify all 10 influenza A virus proteins, including low-abundance proteins like the matrix protein 2 and nonstructural protein 1, with a similar impurity level of host cell proteins. Compared to the centrifugation method, use of the novel magnetic sulfated cellulose particles reduced the influenza A virus particle purification time from 3.5 h to 30 min before mass spectrometry analysis.

Comprehensive N‐glycosylation analysis of the influenza A virus proteins HA and NA from adherent and suspension MDCK cells

“…Cell lines, cell cultivation, and virus infection used in this work were adapted from previous work [20,21]. Briefly, MDCK.ADH cells (ECACC, No.…”

“…Besides others, our group has established several adherent and suspension MDCK cell lines [13,16,17], evaluated virus replication dynamics, and optimized production processes in various bioreactor systems (batch, fed‐batch, plug flow, and hollow fiber) [16,18,19]. Furthermore, we have conducted comprehensive analytical studies including proteomics [20–22] and capillary gel electrophoresis‐based glycoprofiling [23–25]. So far, however, there has not been a detailed investigation on the glycan pattern of the viral envelope proteins HA and NA propagated in our aforementioned MDCK cell lines regarding alterations of N ‐glycan micro‐ and macroheterogeneity.…”

Cellulose as a Scaffold for Self-Assembly: From Basic Research to Real Applications

“…81,82 Serve et al presented a simple method for the purification of influenza virus particles using a novel magnetic sulfated cellulose particle. 83 Compared to the previously established centrifugation method, use of the novel magnetic sulfated cellulose particles reduced the influenza A virus particle purification time from 3.5 h to 30 min before mass spectrometry analysis.…”

“…The electrostatic interaction between the cellulose nanocrystals and chitosan and that between chitosan and Fe 3 O 4 were found to be the key driving forces for the formation of the composite. The obtained magnetic cellulose composites were demonstrated be to potential carriers for the immobilization of enzyme, papain, and pseudomonas cepacialipase. , Serve et al presented a simple method for the purification of influenza virus particles using a novel magnetic sulfated cellulose particle . Compared to the previously established centrifugation method, use of the novel magnetic sulfated cellulose particles reduced the influenza A virus particle purification time from 3.5 h to 30 min before mass spectrometry analysis.…”