PMID- 37400485
OWN - NLM
STAT- MEDLINE
DCOM- 20230705
LR  - 20230706
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 13
IP  - 1
DP  - 2023 Jul 3
TI  - Determining buffer conditions for downstream processing of VLP-based recombinant 
      hepatitis B surface antigen using multimodal resins in bind-elute and 
      flow-through purification modes.
PG  - 10745
LID - 10.1038/s41598-023-37614-y [doi]
LID - 10745
AB  - The difficulties in purification of VLP-based recombinant hepatitis B surface 
      antigen (rHBsAg) are mainly emerged from inefficient semi-purification step plus 
      proteins physicochemical properties and these issues make the downstream 
      processing (DSP) very lengthy and expensive. In this study, optimization of 
      rHBsAg (recombinantly-expressed in Pichia pastoris) DSP was performed using 
      selection of buffering conditions in the semi-purification step. In the 
      semi-purification optimization step, up to 73% of the protein impurities were 
      eliminated and the utmost increase in rHBsAg purity (ca. 3.6-fold) was achieved 
      using 20 mM sodium acetate, pH 4.5. By using rHBsAg binding and nonbinding 
      situations obtained from the response surface plot in design of experiments 
      (DOE), additional bind-elute and flow-through purification mode experiments were 
      conducted and rHBsAg with high purity (near 100%) and recovery (> 83%) was 
      achieved. Following assessment of critical quality attributes (i.e., purity, 
      particle size distribution, host cell DNA, host cell protein, secondary 
      structures, specific activity and relative potency), it was indicated that the 
      characteristics of rHBsAg purified by the new DSP were similar or superior to the 
      ones obtained from conventional DSP. The purification performance of the resin 
      was constantly retained (97-100%) and no significant resin damage took place 
      after 10 adsorption-elution-cleaning cycles. The new DSP developed for production 
      of rHBsAg in this study can substitute the conventional one with granting 
      satisfactory target protein quality, long-lasting resin efficacy, shorter and 
      less expensive process. This process may be also employable for purification of 
      both non-VLP- and VLP- based target proteins expressed in the yeast.
CI  - (c) 2023. The Author(s).
FAU - Goodarzi, Maryam Moazami
AU  - Goodarzi MM
AD  - Department of Research and Development, Production and Research Complex, Pasteur 
      Institute of Iran, Karaj, 3159915111, Iran.
FAU - Jalalirad, Reza
AU  - Jalalirad R
AUID- ORCID: 0000-0003-1826-2570
AD  - Department of Research and Development, Production and Research Complex, Pasteur 
      Institute of Iran, Karaj, 3159915111, Iran. r_jalalirad@pasteur.ac.ir.
FAU - Doroud, Delaram
AU  - Doroud D
AUID- ORCID: 0000-0003-4167-9921
AD  - Department of Research and Development, Production and Research Complex, Pasteur 
      Institute of Iran, Karaj, 3159915111, Iran. d_doroud@pasteur.ac.ir.
FAU - Hozouri, Hamidreza
AU  - Hozouri H
AD  - Department of Quality Management, Production and Research Complex, Pasteur 
      Institute of Iran, Karaj, 3159915111, Iran.
FAU - Aghasadeghi, Mohammadreza
AU  - Aghasadeghi M
AD  - Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, 1316943551, 
      Iran.
AD  - Viral Vaccine Research Center, Pasteur Institute of Iran, Tehran, 1316943551, 
      Iran.
FAU - Paryan, Mahdi
AU  - Paryan M
AD  - Department of Research and Development, Production and Research Complex, Pasteur 
      Institute of Iran, Karaj, 3159915111, Iran.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230703
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Hepatitis B Surface Antigens)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - *Hepatitis B Surface Antigens
MH  - Recombinant Proteins/chemistry
MH  - *Pichia/genetics/metabolism
PMC - PMC10318023
COIS- The authors declare no competing interests.
EDAT- 2023/07/04 01:05
MHDA- 2023/07/05 06:42
PMCR- 2023/07/03
CRDT- 2023/07/03 23:18
PHST- 2022/12/07 00:00 [received]
PHST- 2023/06/24 00:00 [accepted]
PHST- 2023/07/05 06:42 [medline]
PHST- 2023/07/04 01:05 [pubmed]
PHST- 2023/07/03 23:18 [entrez]
PHST- 2023/07/03 00:00 [pmc-release]
AID - 10.1038/s41598-023-37614-y [pii]
AID - 37614 [pii]
AID - 10.1038/s41598-023-37614-y [doi]
PST - epublish
SO  - Sci Rep. 2023 Jul 3;13(1):10745. doi: 10.1038/s41598-023-37614-y.