A novel application of ion exchange chromatography in recombinant hepatitis B vaccine downstream processing: Improving recombinant HBsAg homogeneity by removing associated aggregates

Kimia, Zeinab; Hosseini, Seyed Nezamedin; Talesh, S. Siamak Ashraf; Khatami, Maryam; Kavianpour, Alireza; Javidanbardan, Amin

doi:10.1016/j.jchromb.2019.03.009

Cited by 28 publications

(17 citation statements)

References 43 publications

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“…The process was followed by the separation of biomass from the supernatant and then biomass was disrupted in a bead mill apparatus. It is not necessary to concern the cell viability in separation process [23][24][25][26][27][28][29][30]. Samples were taken from the fermenter and entered the hydrocyclone.…”

Section: Methodsmentioning

confidence: 99%

Numerical and Experimental Biomass Separation from Fermentation Process by Minihydrocyclones

et al. 2020

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Biomass particle separation is challenging in the pharmaceutical industry because of the smallness and lightness of such particles. Centrifugation is applied conventionally for batch separation. However, imposing multiple wash steps in order to eliminate culture residues and the high risk of extrinsic contamination hamper batch techniques. In this research, minihydrocyclones were introduced to provide a better solution for the mentioned limitations in continuous processes. Numerical separation of methylotrophic yeast from fermentation broth was carried out in three hydrocyclones with different lengths, and the most efficient one was examined experimentally. Three various feed flow rates and seven feed concentrations were evaluated. The concentration of biomass suspension in the product was raised to 20 %.

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Section: Methodsmentioning

confidence: 99%

Numerical and Experimental Biomass Separation from Fermentation Process by Minihydrocyclones

et al. 2020

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“…Ion exchange chromatography is the most widely used and cost-effective method for the purification of recombinant proteins. Cation and anion exchange chromatography (CEX and AEX) remove various types of impurities such as product variants, remaining HCP and DNA, media components, leached Protein A, endotoxins, and viruses (Saraswat et al, 2013;Tripathi, 2016;Kimia et al, 2019). The efficacy of a weak anion exchanger on the isolation of rHBsAg VLPs from aggregated structures was also examined, and it was found to yield a 94-97.5% content of rHBsAg VLPs that were within the acceptable quality level (Kimia et al, 2019).…”

Section: Chromatography Processesmentioning

confidence: 99%

“…Cation and anion exchange chromatography (CEX and AEX) remove various types of impurities such as product variants, remaining HCP and DNA, media components, leached Protein A, endotoxins, and viruses (Saraswat et al, 2013;Tripathi, 2016;Kimia et al, 2019). The efficacy of a weak anion exchanger on the isolation of rHBsAg VLPs from aggregated structures was also examined, and it was found to yield a 94-97.5% content of rHBsAg VLPs that were within the acceptable quality level (Kimia et al, 2019). Purification of rHBsAg derived from yeast crude extract was carried out using an AEX column, resulting in high purity (up to >95%) (Ashourian Moghadam et al, 2019).…”

Section: Chromatography Processesmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

369

243

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Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

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“…For this purpose, the small HBsAg S proteins with 226 amino acids are expressed in P. pastoris strains. During the downstream process, virus‐like particles are formed, which contains lipid and about 100 HBsAg S proteins . Due to the molecular size and very hydrophobic nature of HBsAg, efficient extracellular expression of this antigen in the lower eukaryotic systems has not been reported yet .…”

Section: Introductionmentioning

confidence: 99%

“…During the downstream process, virus‐like particles are formed, which contains lipid and about 100 HBsAg S proteins . Due to the molecular size and very hydrophobic nature of HBsAg, efficient extracellular expression of this antigen in the lower eukaryotic systems has not been reported yet . Conventionally, for reaching a high amount of HBsAg, high cell density fermentation process of recombinant P. pastoris Mut + including four main stages has been performed .…”

Section: Introductionmentioning

confidence: 99%

Accurate and cost‐effective prediction of HBsAg titer in industrial scale fermentation process of recombinant Pichia pastoris by using neural network based soft sensor

Hosseini

Javidanbardan

Khatami

2019

Biotech and App Biochem

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In the current work, the attempt was made to apply best-fitted artificial neural network (ANN) architecture and the respective training process for predicting final titer of hepatitis B surface antigen (HBsAg), produced intracellularly by recombinant Pichia pastoris Mut + in the commercial scale. For this purpose, in large-scale fed-batch fermentation, using methanol for HBsAg induction and cell growth, three parameters of average specific growth rate, biomass yield, and dry biomass concentration-in the definite integral form with respect to fermentation time-were selected as input vectors; the final concentration of HBsAg was selected for the ANN output. Used dataset consists of 38 runs from previous batches; feed-forward ANN 3:5:1 with training algorithm of backpropagation based on a Bayesian regularization was trained and tested with a high degree of accuracy. Implementing the verified ANN for predicting the HBsAg titer of the five new fermentation runs, excluded from the dataset, in the full-scale production, the coefficient of regression and root-mean-square error were found to be 0.969299 and 2.716774, respectively. These results suggest that this verified soft sensor could be an excellent alternative for the current relatively expensive and time-intensive analytical techniques such as enzyme-linked immunosorbent assay in the biopharmaceutical industry.

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A novel application of ion exchange chromatography in recombinant hepatitis B vaccine downstream processing: Improving recombinant HBsAg homogeneity by removing associated aggregates

Cited by 28 publications

References 43 publications

Numerical and Experimental Biomass Separation from Fermentation Process by Minihydrocyclones

Numerical and Experimental Biomass Separation from Fermentation Process by Minihydrocyclones

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Accurate and cost‐effective prediction of HBsAg titer in industrial scale fermentation process of recombinant Pichia pastoris by using neural network based soft sensor

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