PMID- 29205941
OWN - NLM
STAT- MEDLINE
DCOM- 20180730
LR  - 20180730
IS  - 1470-8744 (Electronic)
IS  - 0885-4513 (Linking)
VI  - 65
IP  - 3
DP  - 2018 May
TI  - Application of aqueous two-phase micellar system to improve extraction of 
      adenoviral particles from cell lysate.
PG  - 381-389
LID - 10.1002/bab.1627 [doi]
AB  - Viral vectors are important in medical approaches, such as disease prevention and 
      gene therapy, and their production depends on efficient prepurification steps. In 
      the present study, an aqueous two-phase micellar system (ATPMS) was evaluated to 
      extract human adenovirus type 5 particles from a cell lysate. Adenovirus was 
      cultured in human embryonic kidney 293 (HEK-293) cells to a concentration of 1.4 
      x 10(10) particles/mL. Cells were lysed, and the system formed by direct addition 
      of Triton X-114 in a 2(3) full factorial design with center points. The systems 
      were formed with Triton X-114 at a final concentration of 1.0, 6.0, and 11.0% 
      (w/w), cell lysate pH of 6.0, 6.5, and 7.0, and incubation temperatures at 33, 
      35, and 37  degrees C. Adenovirus particles recovered from partition phases were measured 
      by qPCR. The best system condition was with 11.0% (w/w) of Triton X-114, a cell 
      lysate pH of 7.0, and an incubation temperature at 33  degrees C, yielding 3.51 x 10(10) 
      adenovirus particles/mL, which increased the initial adenovirus particles 
      concentration by 2.3-fold, purifying it by 2.2-fold from the cell lysate, and 
      removing cell debris. In conclusion, these results demonstrated that the use of 
      an aqueous two-phase micellar system in the early steps of downstream processing 
      could improve viral particle extraction from cultured cells while integrating 
      clarification, concentration, and prepurification steps.
CI  - (c) 2017 International Union of Biochemistry and Molecular Biology, Inc.
FAU - Molino, Joao Vitor Dutra
AU  - Molino JVD
AUID- ORCID: 0000-0003-2475-9807
AD  - Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical 
      Sciences, University of Sao Paulo, Sao Paulo, Brazil.
FAU - Lopes, Andre Moreni
AU  - Lopes AM
AD  - Department of Bioprocess and Biotechnology, School of Pharmaceutical Sciences, 
      Sao Paulo State University, Araraquara, Brazil.
FAU - Viana Marques, Daniela de Araujo
AU  - Viana Marques DA
AD  - University of Pernambuco, Campus Serra Talhada, Serra Talhada, Brazil.
FAU - Mazzola, Priscila Gava
AU  - Mazzola PG
AD  - Faculty of Pharmaceutical Sciences, University of Campinas, Campinas, Brazil.
FAU - da Silva, Joas Lucas
AU  - da Silva JL
AD  - Department of Clinical and Toxicological Analysis, School of Pharmaceutical 
      Sciences, University of Sao Paulo, Sao Paulo, Brazil.
FAU - Hirata, Mario Hiroyuki
AU  - Hirata MH
AD  - Department of Clinical and Toxicological Analysis, School of Pharmaceutical 
      Sciences, University of Sao Paulo, Sao Paulo, Brazil.
FAU - Hirata, Rosario Dominguez Crespo
AU  - Hirata RDC
AD  - Department of Clinical and Toxicological Analysis, School of Pharmaceutical 
      Sciences, University of Sao Paulo, Sao Paulo, Brazil.
FAU - Gatti, Maria Silvia Viccari
AU  - Gatti MSV
AD  - Department of Genetics, Evolution and Bioagents, Institute of Biology, University 
      of Campinas, Campinas, Brazil.
FAU - Pessoa, Adalberto
AU  - Pessoa A
AD  - Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical 
      Sciences, University of Sao Paulo, Sao Paulo, Brazil.
LA  - eng
PT  - Journal Article
DEP - 20171222
PL  - United States
TA  - Biotechnol Appl Biochem
JT  - Biotechnology and applied biochemistry
JID - 8609465
RN  - 0 (Cell Extracts)
RN  - 0 (Micelles)
RN  - 059QF0KO0R (Water)
SB  - IM
MH  - Adenoviridae/*isolation & purification
MH  - Cell Extracts/*chemistry
MH  - Cells, Cultured
MH  - Genetic Vectors/isolation & purification
MH  - HEK293 Cells
MH  - Humans
MH  - *Micelles
MH  - Water/*chemistry
OTO - NOTNLM
OT  - adenovirus particles
OT  - aqueous two-phase micellar system
OT  - clarification
OT  - partitioning of viral particles
OT  - preconcentration
EDAT- 2017/12/06 06:00
MHDA- 2018/07/31 06:00
CRDT- 2017/12/06 06:00
PHST- 2017/08/03 00:00 [received]
PHST- 2017/11/11 00:00 [accepted]
PHST- 2017/12/06 06:00 [pubmed]
PHST- 2018/07/31 06:00 [medline]
PHST- 2017/12/06 06:00 [entrez]
AID - 10.1002/bab.1627 [doi]
PST - ppublish
SO  - Biotechnol Appl Biochem. 2018 May;65(3):381-389. doi: 10.1002/bab.1627. Epub 2017 
      Dec 22.