Developing approaches, methods, and techniques to make biotherapeutics cheaper and safer.
Bioprocess Intensification
Bioprocess intensification refers to producing more biologics per unit of time, cost, footprint, and volume. Significant intensification can be achieved by both (i) small specific changes to the current development and manufacturing procedures and processes and (ii) more considerable disruptive changes that incorporate developing and implementing novel technologies.
- “Single-Use Centrifugal Separator Enables Intensification of the Clarification Process in Biomanufacturing of Recombinant Proteins.” In review.
Biopharma 4.0
The biopharmaceutical industry is undergoing a digital transformation by incorporating tools and techniques that enable efficient data-driven decision-making across various biopharmaceutical discovery, development, and manufacturing activities. By adopting 4.0 technologies, biopharma companies can accelerate drug discovery and development, increase productivity and enhance regulatory compliance through effective process control, ultimately reducing costs and time associated with biologics development and manufacturing.
- “Artificial neural network (ANN)‐based prediction of depth filter loading capacity for filter sizing.” Biotechnology Progress 32.6 (2016): 1436-1443.
- “Application of Machine Learning in Ensuring Viral Safety of Biotherapeutics: Case Study Demonstrating Prediction and Optimization of Viral Clearance Performance of Anion Exchange Chromatography.” In review.
Continuous Bioprocessing
Switching from batch manufacturing to continuous enables a significant reduction in footprint, ultimately resulting in lower manufacturing costs. Continuous manufacturing processes can be scaled up quickly, based on the operation time, compared to batch processes that require volume-based scale-up. The smaller footprint, lower costs, and faster scale-up of continuous biomanufacturing enable biotherapeutics to be produced at a ‘pandemic pace’ and be accessible to low-income countries.
- “Continuous processing for production of biopharmaceuticals.” Preparative Biochemistry and Biotechnology 45.8 (2015): 836-849.
- “Integrated continuous processing of proteins expressed as inclusion bodies: GCSF as a case study.” Biotechnology progress 33.4 (2017): 998-1009
Publications and Patents
2022
Lynn, David M; Agarwal, Harshit
Liquid Crystal-Infused Slippery Anti-Fouling Surfaces Patent
2022.
@patent{nokey,
title = {Liquid Crystal-Infused Slippery Anti-Fouling Surfaces},
author = {David M Lynn and Harshit Agarwal},
url = {https://patents.google.com/patent/US20220332954A1/en},
year = {2022},
date = {2022-10-20},
abstract = {The present invention provides liquid crystal (LC)-infused materials and methods for detecting compounds or impurities in liquid samples using such materials. These slippery materials comprise a lubricating liquid, preferably a thermotropic liquid crystal, and a solid substrate able to immobilize or host the lubricating liquid. The portion of the substrate coated by the lubricating fluid forms a slippery surface able to allow droplets of various materials to slide off the slippery surface in a manner dependent on the chemical composition of the droplet, which can be used to detect the presence of analytes, impurities and other molecules within the droplet.},
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The present invention provides liquid crystal (LC)-infused materials and methods for detecting compounds or impurities in liquid samples using such materials. These slippery materials comprise a lubricating liquid, preferably a thermotropic liquid crystal, and a solid substrate able to immobilize or host the lubricating liquid. The portion of the substrate coated by the lubricating fluid forms a slippery surface able to allow droplets of various materials to slide off the slippery surface in a manner dependent on the chemical composition of the droplet, which can be used to detect the presence of analytes, impurities and other molecules within the droplet.
Agarwal, Harshit; Quinn, La'Darious J; Walter, Sahana C; Polaske, Thomas J; Chang, Douglas H; Palecek, Sean P; Blackwell, Helen E; Lynn, David M
Slippery Antifouling Polymer Coatings Fabricated Entirely from Biodegradable and Biocompatible Components Journal Article
In: ACS Appl Mater Interfaces, vol. 14, no. 15, pp. 17940–17949, 2022, ISSN: 1944-8252.
@article{pmid35394750,
title = {Slippery Antifouling Polymer Coatings Fabricated Entirely from Biodegradable and Biocompatible Components},
author = {Harshit Agarwal and La'Darious J Quinn and Sahana C Walter and Thomas J Polaske and Douglas H Chang and Sean P Palecek and Helen E Blackwell and David M Lynn},
doi = {10.1021/acsami.1c25218},
issn = {1944-8252},
year = {2022},
date = {2022-04-01},
journal = {ACS Appl Mater Interfaces},
volume = {14},
number = {15},
pages = {17940--17949},
abstract = {We report the design of slippery liquid-infused porous surfaces (SLIPS) fabricated from building blocks that are biodegradable, edible, or generally regarded to be biocompatible. Our approach involves infusion of lubricating oils, including food oils, into nanofiber-based mats fabricated by electrospinning or blow spinning of poly(ε-caprolactone), a hydrophobic biodegradable polymer used widely in medical implants and drug delivery devices. This approach leads to durable and biodegradable SLIPS that prevent fouling by liquids and other materials, including microbial pathogens, on objects of arbitrary shape, size, and topography. This degradable polymer approach also provides practical means to design "controlled-release" SLIPS that release molecular cargo at rates that can be manipulated by the properties of the infused oils (e.g., viscosity or chemical structure). Together, our results provide new designs and introduce useful properties and behaviors to antifouling SLIPS, address important issues related to biocompatibility and environmental persistence, and thus advance new potential applications, including the use of slippery materials for food packaging, industrial and marine coatings, and biomedical implants.},
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We report the design of slippery liquid-infused porous surfaces (SLIPS) fabricated from building blocks that are biodegradable, edible, or generally regarded to be biocompatible. Our approach involves infusion of lubricating oils, including food oils, into nanofiber-based mats fabricated by electrospinning or blow spinning of poly(ε-caprolactone), a hydrophobic biodegradable polymer used widely in medical implants and drug delivery devices. This approach leads to durable and biodegradable SLIPS that prevent fouling by liquids and other materials, including microbial pathogens, on objects of arbitrary shape, size, and topography. This degradable polymer approach also provides practical means to design "controlled-release" SLIPS that release molecular cargo at rates that can be manipulated by the properties of the infused oils (e.g., viscosity or chemical structure). Together, our results provide new designs and introduce useful properties and behaviors to antifouling SLIPS, address important issues related to biocompatibility and environmental persistence, and thus advance new potential applications, including the use of slippery materials for food packaging, industrial and marine coatings, and biomedical implants.
2021
Agarwal, Harshit; Nyffeler, Kayleigh E; Blackwell, Helen E; Lynn, David M
Fabrication of Slippery Liquid-Infused Coatings in Flexible Narrow-Bore Tubing Journal Article
In: ACS Appl Mater Interfaces, vol. 13, no. 46, pp. 55621–55632, 2021, ISSN: 1944-8252.
@article{pmid34775755,
title = {Fabrication of Slippery Liquid-Infused Coatings in Flexible Narrow-Bore Tubing},
author = {Harshit Agarwal and Kayleigh E Nyffeler and Helen E Blackwell and David M Lynn},
doi = {10.1021/acsami.1c14662},
issn = {1944-8252},
year = {2021},
date = {2021-11-01},
journal = {ACS Appl Mater Interfaces},
volume = {13},
number = {46},
pages = {55621--55632},
abstract = {We report a layer-by-layer suction-and-flow approach that enables the fabrication of polymer-based "slippery" liquid-infused porous surfaces (SLIPS) in the confined luminal spaces of flexible, narrow-bore tubing. These SLIPS-coated tubes can prevent or strongly reduce surface fouling after prolonged contact, storage, or flow of a broad range of complex fluids and viscoelastic materials, including many that are relevant in the contexts of medical devices (e.g., blood and urine), food processing (beverages and fluids), and other commercial and industrial applications. The robust and mechanically compliant nature of the nanoporous coating used to host the lubricating oil phase allows these coated tubes to be bent, flexed, and coiled repeatedly without affecting their inherent slippery and antifouling behaviors. Our results also show that SLIPS-coated tubes can prevent the formation of bacterial biofilms after prolonged and repeated flow-based exposure to the human pathogen and that the anti-biofouling properties of these coated tubes can be further improved or prolonged by coupling this approach with strategies that permit the sustained release of broad-spectrum antimicrobial agents. The suction-and-flow approach used here enables the application of slippery coatings in the confined luminal spaces of narrow-bore tubing that are difficult to access using several other methods for the fabrication of liquid-infused coatings and can be applied to tubing of arbitrary length and diameter. We anticipate that the materials and approaches reported here will prove useful for reducing or preventing biofouling, process fouling, and the clogging or occlusion of tubing in a wide range of consumer, industrial, and healthcare-oriented applications.},
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We report a layer-by-layer suction-and-flow approach that enables the fabrication of polymer-based "slippery" liquid-infused porous surfaces (SLIPS) in the confined luminal spaces of flexible, narrow-bore tubing. These SLIPS-coated tubes can prevent or strongly reduce surface fouling after prolonged contact, storage, or flow of a broad range of complex fluids and viscoelastic materials, including many that are relevant in the contexts of medical devices (e.g., blood and urine), food processing (beverages and fluids), and other commercial and industrial applications. The robust and mechanically compliant nature of the nanoporous coating used to host the lubricating oil phase allows these coated tubes to be bent, flexed, and coiled repeatedly without affecting their inherent slippery and antifouling behaviors. Our results also show that SLIPS-coated tubes can prevent the formation of bacterial biofilms after prolonged and repeated flow-based exposure to the human pathogen and that the anti-biofouling properties of these coated tubes can be further improved or prolonged by coupling this approach with strategies that permit the sustained release of broad-spectrum antimicrobial agents. The suction-and-flow approach used here enables the application of slippery coatings in the confined luminal spaces of narrow-bore tubing that are difficult to access using several other methods for the fabrication of liquid-infused coatings and can be applied to tubing of arbitrary length and diameter. We anticipate that the materials and approaches reported here will prove useful for reducing or preventing biofouling, process fouling, and the clogging or occlusion of tubing in a wide range of consumer, industrial, and healthcare-oriented applications.
Agarwal, Harshit; Polaske, Thomas J; Sánchez-Velázquez, Gabriel; Blackwell, Helen E; Lynn, David M
Slippery nanoemulsion-infused porous surfaces (SNIPS): anti-fouling coatings that can host and sustain the release of water-soluble agents Journal Article
In: Chem Commun (Camb), vol. 57, no. 94, pp. 12691–12694, 2021, ISSN: 1364-548X.
@article{pmid34781330,
title = {Slippery nanoemulsion-infused porous surfaces (SNIPS): anti-fouling coatings that can host and sustain the release of water-soluble agents},
author = {Harshit Agarwal and Thomas J Polaske and Gabriel Sánchez-Velázquez and Helen E Blackwell and David M Lynn},
doi = {10.1039/d1cc04645d},
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year = {2021},
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abstract = {We report the design of 'slippery' nanoemulsion-infused porous surfaces (SNIPS). These materials are strongly anti-fouling to a broad range of substances, including microorganisms. Infusion with water-in-oil nanoemulsions also endows these slippery coatings with the ability to host and control or sustain the release of water-soluble agents, including polymers, peptides, and nucleic acids, opening the door to new applications of liquid-infused materials.},
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We report the design of 'slippery' nanoemulsion-infused porous surfaces (SNIPS). These materials are strongly anti-fouling to a broad range of substances, including microorganisms. Infusion with water-in-oil nanoemulsions also endows these slippery coatings with the ability to host and control or sustain the release of water-soluble agents, including polymers, peptides, and nucleic acids, opening the door to new applications of liquid-infused materials.
Agarwal, Harshit; Nyffeler, Kayleigh E; Manna, Uttam; Blackwell, Helen E; Lynn, David M
In: ACS Appl Mater Interfaces, vol. 13, no. 28, pp. 33652–33663, 2021, ISSN: 1944-8252.
@article{pmid34236833,
title = {Liquid Crystal-Infused Porous Polymer Surfaces: A "Slippery" Soft Material Platform for the Naked-Eye Detection and Discrimination of Amphiphilic Species},
author = {Harshit Agarwal and Kayleigh E Nyffeler and Uttam Manna and Helen E Blackwell and David M Lynn},
doi = {10.1021/acsami.1c08170},
issn = {1944-8252},
year = {2021},
date = {2021-07-01},
journal = {ACS Appl Mater Interfaces},
volume = {13},
number = {28},
pages = {33652--33663},
abstract = {We report the design and characterization of liquid crystal (LC)-infused porous polymer membranes that can detect and report on the presence of natural and synthetic amphiphiles in aqueous solution. We demonstrate that thermotropic LCs can be infused into nanoporous polymer membranes to yield LC-infused surfaces that exhibit slippery behaviors in contact with a range of aqueous fluids. In contrast to conventional liquid-infused surfaces (LIS) or slippery liquid-infused porous surfaces (SLIPS) prepared using isotropic oils, aqueous solutions slide over the surfaces of these LC-infused materials at speeds that depend strongly upon the composition of the fluid, including the presence, concentration, or structure of a dissolved surfactant. In general, the sliding times of aqueous droplets on these LC-infused surfaces increase significantly (e.g., from times on the order of seconds to times on the order of minutes) with increasing amphiphile concentration, allowing sliding times to be used to estimate the concentration of the amphiphile. Additional experiments revealed other intrinsic and extrinsic variables or parameters that can be used to further manipulate droplet sliding times and discriminate among amphiphiles of similar structure. Our results are consistent with a physical picture that involves reversible changes in the interfacial orientation of anisotropic LCs mediated by the interfacial adsorption of amphiphiles. These materials thus permit facile "naked-eye" detection and discrimination of amphiphiles in aqueous samples using equipment no more sophisticated than a stopwatch. We demonstrate the potential utility of these LC-infused surfaces for the unaided, naked-eye detection and monitoring of amphiphilic biotoxins in small droplets of fluid extracted directly from cultures of two common bacterial pathogens ( and ). The ability to translate molecular interactions at aqueous/LC interfaces into large and readily observed changes in the sliding times of small aqueous droplets on surfaces could open the door to new applications for antifouling, liquid-infused materials in the context of environmental sensing and other fundamental and applied areas.},
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We report the design and characterization of liquid crystal (LC)-infused porous polymer membranes that can detect and report on the presence of natural and synthetic amphiphiles in aqueous solution. We demonstrate that thermotropic LCs can be infused into nanoporous polymer membranes to yield LC-infused surfaces that exhibit slippery behaviors in contact with a range of aqueous fluids. In contrast to conventional liquid-infused surfaces (LIS) or slippery liquid-infused porous surfaces (SLIPS) prepared using isotropic oils, aqueous solutions slide over the surfaces of these LC-infused materials at speeds that depend strongly upon the composition of the fluid, including the presence, concentration, or structure of a dissolved surfactant. In general, the sliding times of aqueous droplets on these LC-infused surfaces increase significantly (e.g., from times on the order of seconds to times on the order of minutes) with increasing amphiphile concentration, allowing sliding times to be used to estimate the concentration of the amphiphile. Additional experiments revealed other intrinsic and extrinsic variables or parameters that can be used to further manipulate droplet sliding times and discriminate among amphiphiles of similar structure. Our results are consistent with a physical picture that involves reversible changes in the interfacial orientation of anisotropic LCs mediated by the interfacial adsorption of amphiphiles. These materials thus permit facile "naked-eye" detection and discrimination of amphiphiles in aqueous samples using equipment no more sophisticated than a stopwatch. We demonstrate the potential utility of these LC-infused surfaces for the unaided, naked-eye detection and monitoring of amphiphilic biotoxins in small droplets of fluid extracted directly from cultures of two common bacterial pathogens ( and ). The ability to translate molecular interactions at aqueous/LC interfaces into large and readily observed changes in the sliding times of small aqueous droplets on surfaces could open the door to new applications for antifouling, liquid-infused materials in the context of environmental sensing and other fundamental and applied areas.
2017
Kateja, Nikhil; Agarwal, Harshit; Hebbi, Vishwanath; Rathore, Anurag S
Integrated continuous processing of proteins expressed as inclusion bodies: GCSF as a case study Journal Article
In: Biotechnol Prog, vol. 33, no. 4, pp. 998–1009, 2017, ISSN: 1520-6033.
@article{pmid27977908,
title = {Integrated continuous processing of proteins expressed as inclusion bodies: GCSF as a case study},
author = {Nikhil Kateja and Harshit Agarwal and Vishwanath Hebbi and Anurag S Rathore},
doi = {10.1002/btpr.2413},
issn = {1520-6033},
year = {2017},
date = {2017-07-01},
journal = {Biotechnol Prog},
volume = {33},
number = {4},
pages = {998--1009},
abstract = {Affordability of biopharmaceuticals continues to be a challenge, particularly in developing economies. This has fuelled advancements in manufacturing that can offer higher productivity and better economics without sacrificing product quality in the form of an integrated continuous manufacturing platform. While platform processes for monoclonal antibodies have existed for more than a decade, development of an integrated continuous manufacturing process for bacterial proteins has received relatively scant attention. In this study, we propose an end-to-end integrated continuous downstream process (from inclusion bodies to unformulated drug substance) for a therapeutic protein expressed in Escherichia coli as inclusion body. The final process consisted of a continuous refolding in a coiled flow inverter reactor directly coupled to a three-column periodic counter-current chromatography for capture of the product followed by a three-column con-current chromatography for polishing. The continuous bioprocessing train was run uninterrupted for 26 h to demonstrate its capability and the resulting output was analyzed for the various critical quality attributes, namely product purity (>99%), high molecular weight impurities (<0.5%), host cell proteins (<100 ppm), and host cell DNA (<10 ppb). All attributes were found to be consistent over the period of operation. The developed assembly offers smaller facility footprint, higher productivity, fewer hold steps, and significantly higher equipment and resin utilization. The complexities of process integration in the context of continuous processing have been highlighted. We hope that the study presented here will promote development of highly efficient, universal, end-to-end, fully continuous platforms for manufacturing of biotherapeutics. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:998-1009, 2017.},
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}
Affordability of biopharmaceuticals continues to be a challenge, particularly in developing economies. This has fuelled advancements in manufacturing that can offer higher productivity and better economics without sacrificing product quality in the form of an integrated continuous manufacturing platform. While platform processes for monoclonal antibodies have existed for more than a decade, development of an integrated continuous manufacturing process for bacterial proteins has received relatively scant attention. In this study, we propose an end-to-end integrated continuous downstream process (from inclusion bodies to unformulated drug substance) for a therapeutic protein expressed in Escherichia coli as inclusion body. The final process consisted of a continuous refolding in a coiled flow inverter reactor directly coupled to a three-column periodic counter-current chromatography for capture of the product followed by a three-column con-current chromatography for polishing. The continuous bioprocessing train was run uninterrupted for 26 h to demonstrate its capability and the resulting output was analyzed for the various critical quality attributes, namely product purity (>99%), high molecular weight impurities (<0.5%), host cell proteins (<100 ppm), and host cell DNA (<10 ppb). All attributes were found to be consistent over the period of operation. The developed assembly offers smaller facility footprint, higher productivity, fewer hold steps, and significantly higher equipment and resin utilization. The complexities of process integration in the context of continuous processing have been highlighted. We hope that the study presented here will promote development of highly efficient, universal, end-to-end, fully continuous platforms for manufacturing of biotherapeutics. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:998-1009, 2017.
2016
Agarwal, Harshit; Rathore, Anurag S; Hadpe, Sandeep Ramesh; Alva, Solomon J
Artificial neural network (ANN)-based prediction of depth filter loading capacity for filter sizing Journal Article
In: Biotechnol Prog, vol. 32, no. 6, pp. 1436–1443, 2016, ISSN: 1520-6033.
@article{pmid27453285,
title = {Artificial neural network (ANN)-based prediction of depth filter loading capacity for filter sizing},
author = {Harshit Agarwal and Anurag S Rathore and Sandeep Ramesh Hadpe and Solomon J Alva},
doi = {10.1002/btpr.2329},
issn = {1520-6033},
year = {2016},
date = {2016-11-01},
journal = {Biotechnol Prog},
volume = {32},
number = {6},
pages = {1436--1443},
abstract = {This article presents an application of artificial neural network (ANN) modelling towards prediction of depth filter loading capacity for clarification of a monoclonal antibody (mAb) product during commercial manufacturing. The effect of operating parameters on filter loading capacity was evaluated based on the analysis of change in the differential pressure (DP) as a function of time. The proposed ANN model uses inlet stream properties (feed turbidity, feed cell count, feed cell viability), flux, and time to predict the corresponding DP. The ANN contained a single output layer with ten neurons in hidden layer and employed a sigmoidal activation function. This network was trained with 174 training points, 37 validation points, and 37 test points. Further, a pressure cut-off of 1.1 bar was used for sizing the filter area required under each operating condition. The modelling results showed that there was excellent agreement between the predicted and experimental data with a regression coefficient (R ) of 0.98. The developed ANN model was used for performing variable depth filter sizing for different clarification lots. Monte-Carlo simulation was performed to estimate the cost savings by using different filter areas for different clarification lots rather than using the same filter area. A 10% saving in cost of goods was obtained for this operation. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1436-1443, 2016.},
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This article presents an application of artificial neural network (ANN) modelling towards prediction of depth filter loading capacity for clarification of a monoclonal antibody (mAb) product during commercial manufacturing. The effect of operating parameters on filter loading capacity was evaluated based on the analysis of change in the differential pressure (DP) as a function of time. The proposed ANN model uses inlet stream properties (feed turbidity, feed cell count, feed cell viability), flux, and time to predict the corresponding DP. The ANN contained a single output layer with ten neurons in hidden layer and employed a sigmoidal activation function. This network was trained with 174 training points, 37 validation points, and 37 test points. Further, a pressure cut-off of 1.1 bar was used for sizing the filter area required under each operating condition. The modelling results showed that there was excellent agreement between the predicted and experimental data with a regression coefficient (R ) of 0.98. The developed ANN model was used for performing variable depth filter sizing for different clarification lots. Monte-Carlo simulation was performed to estimate the cost savings by using different filter areas for different clarification lots rather than using the same filter area. A 10% saving in cost of goods was obtained for this operation. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1436-1443, 2016.
Kateja, Nikhil; Agarwal, Harshit; Saraswat, Aditya; Bhat, Manish; Rathore, Anurag S
In: Biotechnol J, vol. 11, no. 10, pp. 1320–1331, 2016, ISSN: 1860-7314.
@article{pmid27447837,
title = {Continuous precipitation of process related impurities from clarified cell culture supernatant using a novel coiled flow inversion reactor (CFIR)},
author = {Nikhil Kateja and Harshit Agarwal and Aditya Saraswat and Manish Bhat and Anurag S Rathore},
doi = {10.1002/biot.201600271},
issn = {1860-7314},
year = {2016},
date = {2016-10-01},
journal = {Biotechnol J},
volume = {11},
number = {10},
pages = {1320--1331},
abstract = {Coiled Flow Inverter Reactor (CFIR) has recently been explored for facilitating continuous operation of several unit operations involved in downstream processing of biopharmaceuticals such as viral inactivation and protein refolding. The application of CFIR for continuous precipitation of clarified cell culture supernatant has been explored. The pH based precipitation is optimized in the batch mode and then in the continuous mode in CFIR using a design of experiments (DOE) study. Improved clearance of host cell DNA (52× vs. 39× in batch), improved clearance of host cell proteins (HCP) (7× vs. 6× in batch) and comparable recovery (90 vs. 91.5 % in batch) are observed along with six times higher productivity. To further demonstrate wider applicability of CFIR in performing continuous precipitation, two more case studies involving use of two different precipitation protocols (CaCl based and caprylic acid based) are also performed. In both cases, clearance of host cell DNA, HCP, and product recovery are found to be comparable or better in CFIR than in batch operations. Moreover, increase in productivity of 16 times (CaCl based) and eight times (caprylic acid based) is obtained for the two precipitation protocols, respectively. The data clearly demonstrate that CFIR can be seamlessly integrated into a continuous bioprocess train for performing continuous precipitation of clarified cell culture supernatant. To our knowledge this is the first report of such use.},
keywords = {},
pubstate = {published},
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Coiled Flow Inverter Reactor (CFIR) has recently been explored for facilitating continuous operation of several unit operations involved in downstream processing of biopharmaceuticals such as viral inactivation and protein refolding. The application of CFIR for continuous precipitation of clarified cell culture supernatant has been explored. The pH based precipitation is optimized in the batch mode and then in the continuous mode in CFIR using a design of experiments (DOE) study. Improved clearance of host cell DNA (52× vs. 39× in batch), improved clearance of host cell proteins (HCP) (7× vs. 6× in batch) and comparable recovery (90 vs. 91.5 % in batch) are observed along with six times higher productivity. To further demonstrate wider applicability of CFIR in performing continuous precipitation, two more case studies involving use of two different precipitation protocols (CaCl based and caprylic acid based) are also performed. In both cases, clearance of host cell DNA, HCP, and product recovery are found to be comparable or better in CFIR than in batch operations. Moreover, increase in productivity of 16 times (CaCl based) and eight times (caprylic acid based) is obtained for the two precipitation protocols, respectively. The data clearly demonstrate that CFIR can be seamlessly integrated into a continuous bioprocess train for performing continuous precipitation of clarified cell culture supernatant. To our knowledge this is the first report of such use.
2015
Rathore, Anurag S; Agarwal, Harshit; Sharma, Abhishek Kumar; Pathak, Mili; Muthukumar, S
Continuous processing for production of biopharmaceuticals Journal Article
In: Prep Biochem Biotechnol, vol. 45, no. 8, pp. 836–849, 2015, ISSN: 1532-2297.
@article{pmid25674930,
title = {Continuous processing for production of biopharmaceuticals},
author = {Anurag S Rathore and Harshit Agarwal and Abhishek Kumar Sharma and Mili Pathak and S Muthukumar},
doi = {10.1080/10826068.2014.985834},
issn = {1532-2297},
year = {2015},
date = {2015-01-01},
journal = {Prep Biochem Biotechnol},
volume = {45},
number = {8},
pages = {836--849},
abstract = {The merits of continuous processing over batch processing are well known in the manufacturing industry. Continuous operation results in shorter process times due to omission of hold steps, higher productivity due to reduced shutdown costs, and lowers labor requirement. Over the past decade, there has been an increasing interest in continuous processing within the bioprocessing community, specifically those involved in production of biotherapeutics. Continuous operations in upstream processing (perfusion) have been performed for decades. However, recent development of continuous downstream operations has led the industry to envisage an integrated bioprocessing platform for efficient production. The regulators, key players in the biotherapeutic industry, have also expressed their interest and willingness in this migration from the traditional batch processing. This paper aims to review major developments in continuous bioprocessing in the past decade. A discussion of pros and cons of the different proposed approaches has also been presented.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The merits of continuous processing over batch processing are well known in the manufacturing industry. Continuous operation results in shorter process times due to omission of hold steps, higher productivity due to reduced shutdown costs, and lowers labor requirement. Over the past decade, there has been an increasing interest in continuous processing within the bioprocessing community, specifically those involved in production of biotherapeutics. Continuous operations in upstream processing (perfusion) have been performed for decades. However, recent development of continuous downstream operations has led the industry to envisage an integrated bioprocessing platform for efficient production. The regulators, key players in the biotherapeutic industry, have also expressed their interest and willingness in this migration from the traditional batch processing. This paper aims to review major developments in continuous bioprocessing in the past decade. A discussion of pros and cons of the different proposed approaches has also been presented.