TY - JOUR
T1 - Viral vector purification with thermoresponsive-anionic mixed polymer brush modified beads-packed column
AU - Nagase, Kenichi
AU - Kitazawa, Sakiko
AU - Kogure, Toshihiko
AU - Yamada, Sota
AU - Katayama, Kazuhiro
AU - Kanazawa, Hideko
N1 - Funding Information:
This work was partly supported financially by a SENTAN grant JPMJSN16B3 from the Japan Science and Technology Agency (JST) and a Grant-in-aid for Scientific Research (grant no. 19H02447 and 20H05233) from the Japan Society for the Promotion of Science (JSPS).
Publisher Copyright:
© 2022 Elsevier B.V.
PY - 2022/4/1
Y1 - 2022/4/1
N2 - Viral vectors have attracted attention as a potential new therapeutic modality for gene therapy. In this study, we developed a temperature-modulated viral vector purification column using a mixed polymer brush composed of thermoresponsive and anionic polymers as packing material ligands. The mixed polymer brushes were modified on silica beads using a combination of reversible addition − fragmentation chain transfer (RAFT) polymerization of 2-acrylamido-2-methylpropanesulfonic acid (AMPS) and subsequent atom transfer radical polymerization (ATRP) of N-isopropylacrylamide (NIPAAm). The temperature-modulated zeta potential change in the prepared mixed polymer brush was attributed to the PNIPAAm shrinking and exposing of PAMPS. The prepared mixed polymer brush modified beads were used as packing materials, and the elution behavior of the adeno associated virus type 2 (AAV2) vector was observed. The AAV2 vector was adsorbed on the mixed polymer brush by electrostatic and hydrophobic interactions at 40 °C. By reducing the temperature to 5 °C, adsorbed AAV2 vector on the mixed polymer brush was desorbed and eluted from the column due to lowering the electrostatic and hydrophobic interactions between the AAV2 vector and mixed polymer brush. The AAV2 vector was separated from bovine serum proteins as a contaminant using the column. The ability to infect cells was maintained by the recovered AAV2 vectors from the column. Thus, the developed column would be beneficial for the simple purification of viral vectors.
AB - Viral vectors have attracted attention as a potential new therapeutic modality for gene therapy. In this study, we developed a temperature-modulated viral vector purification column using a mixed polymer brush composed of thermoresponsive and anionic polymers as packing material ligands. The mixed polymer brushes were modified on silica beads using a combination of reversible addition − fragmentation chain transfer (RAFT) polymerization of 2-acrylamido-2-methylpropanesulfonic acid (AMPS) and subsequent atom transfer radical polymerization (ATRP) of N-isopropylacrylamide (NIPAAm). The temperature-modulated zeta potential change in the prepared mixed polymer brush was attributed to the PNIPAAm shrinking and exposing of PAMPS. The prepared mixed polymer brush modified beads were used as packing materials, and the elution behavior of the adeno associated virus type 2 (AAV2) vector was observed. The AAV2 vector was adsorbed on the mixed polymer brush by electrostatic and hydrophobic interactions at 40 °C. By reducing the temperature to 5 °C, adsorbed AAV2 vector on the mixed polymer brush was desorbed and eluted from the column due to lowering the electrostatic and hydrophobic interactions between the AAV2 vector and mixed polymer brush. The AAV2 vector was separated from bovine serum proteins as a contaminant using the column. The ability to infect cells was maintained by the recovered AAV2 vectors from the column. Thus, the developed column would be beneficial for the simple purification of viral vectors.
KW - Bioseparation
KW - Temperature-responsive chromatography
KW - Thermoresponsive polymer
KW - Viral vector
UR - http://www.scopus.com/inward/record.url?scp=85122498307&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85122498307&partnerID=8YFLogxK
U2 - 10.1016/j.seppur.2022.120445
DO - 10.1016/j.seppur.2022.120445
M3 - Article
AN - SCOPUS:85122498307
VL - 286
JO - Gas Separation and Purification
JF - Gas Separation and Purification
SN - 1383-5866
M1 - 120445
ER -