WO2016116947A1 - Réacteur d'inversion de flux à serpentins pour le repliement continu de protéines recombinantes dénaturées et autres opérations de mélange - Google Patents

Réacteur d'inversion de flux à serpentins pour le repliement continu de protéines recombinantes dénaturées et autres opérations de mélange Download PDF

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Publication number
WO2016116947A1
WO2016116947A1 PCT/IN2016/000022 IN2016000022W WO2016116947A1 WO 2016116947 A1 WO2016116947 A1 WO 2016116947A1 IN 2016000022 W IN2016000022 W IN 2016000022W WO 2016116947 A1 WO2016116947 A1 WO 2016116947A1
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Prior art keywords
reactor
refolding
coiled
flow inverter
protein
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PCT/IN2016/000022
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English (en)
Inventor
Anurag Rathore
Krishna Deo Prasad Nigam
Mili Pathak
Harshit Agarwal
Abhishek Kumar Sharma
Nikhil Kateja
Vishwanath Hebbi
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Indian Institute Of Technology
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Publication of WO2016116947A1 publication Critical patent/WO2016116947A1/fr

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/24—Stationary reactors without moving elements inside
    • B01J19/2415—Tubular reactors
    • B01J19/243—Tubular reactors spirally, concentrically or zigzag wound
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
    • B01F25/40—Static mixers
    • B01F25/42—Static mixers in which the mixing is affected by moving the components jointly in changing directions, e.g. in tubes provided with baffles or obstructions
    • B01F25/43—Mixing tubes, e.g. wherein the material is moved in a radial or partly reversed direction
    • B01F25/433—Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
    • B01F25/4331—Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
    • C07K1/1136—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure by reversible modification of the secondary, tertiary or quarternary structure, e.g. using denaturating or stabilising agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/53—Colony-stimulating factor [CSF]
    • C07K14/535—Granulocyte CSF; Granulocyte-macrophage CSF

Definitions

  • the present invention relates with the designing of a novel reactor. More 10 particularly, the present invention involves the use of a Coiled Flow Inverter Reactor (CFIR) for achieving homogeneity in concentrations of protein and other reactants throughout the process so as to achieve the desirable outcome of the resulting reaction.
  • the reaction could be refolding, precipitation, viral inactivation etc.
  • the reactor consists of an inline mixer 15 followed by a coiled flow inverter that is designed to provide cross sectional mixing of held up protein solution.
  • This invention relates to ensuring successful translation of biotech unit operations that require concentrations of protein and other reactants to be kept homogenous throughout the process so as to achieve the desirable outcome of the resulting reaction.
  • the invention does so by employing a modified design of a coiled flow inverter (CFI) first introduced in U.S. Patent No. 7,337,835, which causes cross sectional mixing of the holdup via flow inversion that is achieved by changing the direction of centrifugal force in helically-coiled tubes.
  • CFI coiled flow inverter
  • CFIR is a configuration of equally spaced helical coils in each bank that are substantially in one common plane, the axis of each helical coil is at an angle of 90° bent to the adjacent helical coil. These bends at right angles cause flow inversion of liquids, thus enhancing cross sectional mixing in the tube.
  • the configuration can be appropriately utilized to provide a sharper residence time distribution along with good cross-sectional mixing.
  • Recombinant proteins are often over expressed in microbial systems such as E.coli, resulting in formation of insoluble inclusion bodies. This necessitates denaturation of the product in these inclusion bodies and requires refolding in order to attain the native structure.
  • the traditional and most widely used industrial process is in batch reactor, wherein denatured protein is diluted in refolding buffer placed in stirred tank (Bade et al, 2012). This often ends in very dilute solutions with large handling of process volumes and is time consuming. This poses the need for designing a continuous refolding reactor that meets the process requirements.
  • CSTR continuous stirred tank reactor
  • CSTR continuous stirred tank reactor
  • U.S. Patent No. 4,999,422 shows that the use of CSTR for continuous refolding provides broad residence time distribution that has been known to lead to misfolded and degraded proteins along with native protein. Further, shear and foaming can cause aggregation and product loss.
  • the other U.S. Patent No. 8,067,201 describes an approach involving dialysis based refolding wherein yield of refolded solution is 90% with 32X dilution. This approach in addition to above mentioned drawbacks increases the volume and restricts process operation from happening in real continuous mode.
  • Schlegl et al (2005) have proposed use of a CSTR along with a diafiltration circuit to lower the denaturant levels.
  • the folded protein is removed selectively and unfolded protein is recycled back. This, however, poses a problem of selective removal of refolded protein.
  • Pan et al (2014) have designed a tubular reactor for continuous refolding of proteins with dilution of 10X and residence time of 5 to 21 hours for different proteins.
  • This tubular reactor works as a PFR with a long run time.
  • a CSTR provides better mixing but is bound to have a broad residence time distribution, whereas a plug flow reactor has a narrower residence time distribution but may lack the necessary mixing required for refolding reaction.
  • Continuous precipitation has been performed in a variety of configurations.
  • Tavare and Patwardhan (1992) have performed crystallization of copper sulphate, nickel ammonium sulfate, and soy protein in continuous mixed suspension and mixed product removal reactors (MSMPR).
  • MSMPR continuous mixed suspension and mixed product removal reactors
  • the product crystal size distributions at steady state was measured and correlated in terms of power law kinetics to understand the effect of various observable variables.
  • An MSMPR precipitator with a volume of 273ml was used by Raphael and Rohani in 1996 for isoelectric precipitation of sunflower protein.
  • Pan et al (2014) have used an integrated continuous tubular reactor system for processing an autoprotease expressed as inclusion bodies.
  • the inclusion bodies were suspended and fed into the tubular reactor system for continuous dissolving, refolding and precipitation.
  • Refolding and precipitation yield in tubular reactor were similar to batch reactor and process was stable for at least 20h.
  • Productivity in mg/l/h was found to be twice in tubular reactor compared to batch reactor.
  • the present invention intends to provide a system for achieving homogeneity in concentrations of protein and other reactants throughout the process so as to achieve the desirable outcome of the resulting reaction.
  • the reaction could be refolding, precipitation, viral inactivation etc.
  • the proposed invention has been successfully used for performing refolding of GCSF.
  • the CFIR has also been successfully used for translating batch precipitation process into a continuous one by continuous mixing of a clarified harvest stream getting homogenized with a stream of acid. Both applications demonstrate the realization of the benefits expected from continuous processing.
  • the main object of the present invention is to provide an effective reactor design, enabling continuous processing of protein dispersions in biotech unit operations.
  • the other object of the present invention is to provide a reactor which enables effective mixing required for biotech unit operations in order to maintain homogenous physicochemical environment for the proteins and other biomolecules to refold to improve contacting in case of a reaction and avoid protein aggregation.
  • Another object of the present invention is to provide an effective reactor design based on coiled flow inverter geometry.
  • Another object of the present invention is to construct a coiled flow inverter reactor in a Single Use Technology paradigm where the entire configuration can be made out of autoclavable materials.
  • Yet another object of the present invention is to provide a reactor designed for refolding which can be configured to work for various proteins by configuring well defined design parameters such as the residence time, flow rates, buffer composition etc.
  • Another object of the present invention is to provide a reactor designed for precipitation of impurities in a cell culture harvest which can be configured to work for various proteins by configuring well defined design parameters such as the residence time, flow rates, buffer composition etc.
  • Another object of the present invention is to provide a versatile reactor configuration for protein processing for a variety of biotech unit operations such asviral inactivation.
  • Still another object of the present invention is that the reactor designed should seamlessly connect as a module with other unit operations before and after within an integrated/ continuous bioprocessing platform.
  • the present invention relates to an innovative reactor (CFIR) system based on coiled flow inverter geometry which consists of a number of banks of tubes, each formed by four discretely wound helical coils which are appreciably coplanar, with each coil having equal number of turns and such that axes of adjacent coils is separated by a right angle before and after 90° bend as shown in Fig. 1.
  • CFIR innovative reactor
  • the reactor continuously processes a stream protein dispersion, possibly mixed with additives during the process keeping it homogenous and stable.
  • the coiled flow inverter geometry imparts secondary flow in the cross-section of the tube that changes direction after each bend, thus causing mixing in a sustained fashion.
  • the mixing flattens the velocity profile such that the residence time distribution and hence, if applicable, the degree of conversion in the process at a given point along the length is narrow and thus the process can be configured to achieve high biologically active protein purity at the outlet for a given protein. This maintains the homogeneity and stability of the protein solution, preventing aggregation and precipitation of the protein, respectively.
  • the reactor processes a continuous stream of denatured protein by diluting it with a buffer that creates an environment conducive for proper refolding in order to provide a stable stream of biologically active protein at the outlet.
  • the reactor is also capable of continuously precipitating out impurities in clarified harvest from a mammalian cell culture based bioreactor by providing a homogeneous precipitation environment to the harvest for narrow residence time distributions.
  • Figure 1 shows the schematic of a 90° bend of a Coiled Flow Inverter (CFI) of one of the embodiment of the invention wherein 5 turns are used in each of the arm.
  • the curvature ratio is 8.4 and the Deans number is 3.27.
  • Figure 2 is a process flow diagram of refolding in CFIR.
  • Figure 3 shows the refolding protocol of recombinant human granulocyte colony stimulating factor(GCSF). It can be seen that the unfolded protein refolds to give rise to a saturated equilibrium level of the percentage native (correctly folded) protein.
  • GCSF granulocyte colony stimulating factor
  • Figure 4 shows comparison of continuous (a) and batch (b) refolding process for recombinant human granulocyte colony stimulating factor (rhGCSF).
  • Figure 5 presents the schematics for the experimental set-up of precipitation process performed using coiled flow inverter reactor.
  • the various embodiments of the present invention provides a reactor design configured for processing of protein dispersions continuously while maintaining the stability of the dispersion.
  • continuous refolding of denatured proteins is accomplished by processing of continuously flowing streams of refolding buffer and reduced recombinant proteins to obtain high productivity of refolded protein at the outlet continuously.
  • the old traditional batch process involves dilution of a protein suspension and requires initial mixing of there folding buffer and the reduced inclusion bodies, followed by sustained mixing of the resulting solution until the native conformation is attained.
  • a refolding process has two critical aspects: initial mixing and process mixing.
  • Initial mixing is critical for process performance (Mannall et al, 2006), as poor mixing can lead to regions of higher intrinsic intermediate concentration that can cause aggregation of proteins.
  • the sustained process mixing is generally required to homogenize the protein solution during the course of refolding to prevent intermolecular interactions among proteins and hence prevent protein aggregation.
  • the use of the coiled flow inverter geometry fulfills the mixing requirement for protein folding reaction.
  • the coiled flow inverter geometry consists of a number of banks, each formed by four discretely wound helical coils which are appreciably coplanar, with each coil having equal number of turns and such that axes of adjacent coils is separated by a right angle as shown in Figure 1. While motion of fluid in a helical path leads to secondary flow in the cross section caused by centrifugal force, bending the tube leads to change in the direction of secondary flow pattern, leading to flow inversion and improved mixing. While mixing, coiled flow inverter geometry homogenizes and stabilizes the protein solution, it also flattens the cross sectional velocity profile, hence better emulating a plug flow due to a narrower residence time distribution.
  • this effect is utilized to get a narrow distribution of degree of refolding at a given cross-section along the tube length.
  • This also implies that there would be no requirement of additional separation and recycling of unfolded protein which otherwise adds to the complications of validation of reprocessing.
  • the outlet protein solution can be readily connected to the next unit operation in a seamless fashion, preventing mix-ups and contamination resulting in improved compliance with the current good manufacturing practices.
  • the present invention is exemplified with reference to recombinant human granulocyte colony stimulating factor (rhGCSF). The experimental setup and process flow has been shown in detail in Figures 2 and 3.
  • a bank is a collection of four branches, each with five turns of helix.
  • refolding buffer and reduced IBs are pumped at defined flow rates to achieve the required dilution ratio through the coiled flow inverter.
  • An inline mixer is added before the CFIR to provide the fast mixing required at the start of refolding.
  • the sample is quenched with acetic acid and stored.
  • two peristaltic pumps were used to pump refolding buffer and reduced inclusion bodies in the desired ratio of 5: 1 .
  • Total 3 banks were used and sampling was done at different time points.
  • a dynamic inline mixture was used at the beginning to provide rapid initial mixing. Quenching of the refolded sample was done using inline dilution by acetic acid as shown in Figure 2
  • Figure 3 explains the refolding protocol occurring inside the invention.
  • the inclusion bodies are first solubilized using solubilization buffer (50 mM Tris buffer pH 10, containing 6 M Urea) by constant mixing on a stirrer at 180 rpm for 45 min at room temperature.
  • the reducing agent Dithiothreitol (DTT)
  • DTT Dithiothreitol
  • the reduced solubilized IBs are diluted in the refolding buffer in the desired ratio of 1 :5 using dynamic inline mixture. This solution is then given the desired residence time in CFIR.
  • the samples are quenched using glacial acetic acid at pH 4 to stop refolding reaction and are subjected for further analysis.
  • the RP-HPLC data for batch refolding and continuous refolding process depicts that it takes 150 minutes for batch process to give 83% native protein (sampling point 30min, 60min, 120min, 150min, 180min, 240min, 480min, 720min and 960min) while 84% native protein is achieved in 18min using CFIR (sampling point Omin, 18min, 30min, 60min, 90min, 120min, 150min and 180min). As per our calculations, productivity of continuous process will be17 times higher than batch process for the application under consideration.
  • percentage of native protein with time of refold shows the native percentage approach 84% after 1 h of refold.
  • Percentage of refolded native GCSF in batch process as a function of time is determined by RP-HPLC. It is seen that the refolding is complete after 1 h 30 min and results in 84% purity.
  • the CFIR enables continuous refolding of proteins with high protein concentration with increased purity and hence provides better productivity as seen in Figure 4. It also easily handles large flow rates and large volumes.
  • the reactor can be easily configured for refolding of different proteins by changing flow rates, number of branches and other process parameters.
  • the reactor can be modified for refolding various other proteins by configuring process parameters such as residence time, buffer composition, flow rate etc.
  • process parameters such as residence time, buffer composition, flow rate etc.
  • the described invention is a compact embodiment of a continuous flow reactor that provides sustained mixing of the held-up volume with minimized back mixing and hence a sharp residence time distribution. Further, the incorporation of the CFIR flow design enables a great degree of customizability to the reactor such that one can vary critical design attributes to optimally customize it to any given process. For example:
  • the variation in Dean number can be performed conveniently by varying the helical radius and/or tube radius to manipulate the degree of mixing.
  • the residence time distribution can be narrowed down conveniently as understood from the geometrical parameters and flow rate in the coiled flow inverter.
  • Possible applications of the invention include a variety of reactions such as protein refolding, viral inactivation, precipitation, two phase separation, etc. Further, the invention enables inline addition and/or modification at any point along the reactor length to of process conditions to incorporate various process strategies. For example:
  • the present invention is amenable to a single-use construction and the sharper residence times make it amenable towards effective control schemes in a continuous bioprocessing train.
  • the reactor can be seen as a modular unit connected seamlessly with an inlet and an outlet in a continuous bioprocessing train.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
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  • Genetics & Genomics (AREA)
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Abstract

La présente invention concerne une nouvelle conception de réacteur basée sur une géométrie d'inversion de flux à serpentins pour la mise en oeuvre de réactions de repliement de protéines en continu in vitro par un procédé de dilution. Le réacteur est composé d'un mélangeur en ligne suivi d'une conception visant à fournir un mélange transversal de protéines dénaturées diluées par inversion de flux ce qui permet de changer la direction de la force centrifuge dans un tube hélicoïdal. La caractéristique unique du réacteur d'inversion de flux à serpentins (CFIR) est qu'il améliore le contact parmi les différentes entités telles que chaotropes, stabilisants, ions tampon, en réduisant et en oxydant des agents et des molécules de protéines, à l'intérieur de la section transversale dans un environnement de faible cisaillement, ce qui permet d'obtenir une distribution pointue des temps de séjour et de stabiliser la réaction de repliement par réduction des réactions secondaires qui autrement entraînent la formation d'agrégats et d'impuretés oxydées et aboutissent à la réduction du rendement du repliement. Le réacteur est également illustré pour sa polyvalence : il est applicable à d'autres opérations unitaires de biotechnologie par démonstration d'une précipitation continue d'impuretés à partir de la récolte de cultures cellulaires d'un anticorps monoclonal thérapeutique. La présente invention peut être utilisée pour obtenir une homogénéité des concentrations de protéines et autres réactifs tout au long du processus de manière à obtenir le résultat attendu de la réaction résultante pour le repliement, la précipitation, l'inactivation virale etc. des protéines.
PCT/IN2016/000022 2015-01-21 2016-01-20 Réacteur d'inversion de flux à serpentins pour le repliement continu de protéines recombinantes dénaturées et autres opérations de mélange WO2016116947A1 (fr)

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107759659A (zh) * 2016-08-19 2018-03-06 昂德生物药业有限公司 注射用重组人组织型纤溶酶原激酶衍生物制备用的复性稀释分散装置及其应用
WO2018138566A1 (fr) * 2017-01-27 2018-08-02 Indian Institute Of Technology, Delhi Onduleurs à flux spiralé compacts utiles en tant que mélangeurs en ligne
WO2020095894A1 (fr) * 2018-11-05 2020-05-14 味の素株式会社 Procédé de production d'une protéine repliée à l'aide d'un microréacteur à flux, et appareil de repliement de protéine
WO2021111470A1 (fr) * 2019-12-03 2021-06-10 Indian Institute Of Technology Delhi Procédé de préparation protéines thérapeutiques pégylées
WO2021230956A1 (fr) * 2020-05-11 2021-11-18 Purdue Research Foundation Synthèse à l'échelle de lomustine dans des conditions d'écoulement continu
EP3984627A1 (fr) 2020-10-19 2022-04-20 Novasep Equipment Solutions Appareil et procédé de purification de biomolécules
CN115003635A (zh) * 2020-02-03 2022-09-02 默克专利股份有限公司 模块化培养室和病毒灭活方法
US11520310B2 (en) 2019-06-18 2022-12-06 International Business Machines Corporation Generating control settings for a chemical reactor
US11675334B2 (en) 2019-06-18 2023-06-13 International Business Machines Corporation Controlling a chemical reactor for the production of polymer compounds

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4999422A (en) 1988-04-15 1991-03-12 Biogen, N.V. Continuous method of refolding proteins
US20060162912A1 (en) * 2005-01-25 2006-07-27 Indian Institute Of Technology Delhi Heat exchanger
US8067201B2 (en) 2009-04-17 2011-11-29 Bristol-Myers Squibb Company Methods for protein refolding
WO2014155349A2 (fr) * 2013-03-29 2014-10-02 Dr. Reddy's Laboratories Repliement de protéines

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4999422A (en) 1988-04-15 1991-03-12 Biogen, N.V. Continuous method of refolding proteins
US20060162912A1 (en) * 2005-01-25 2006-07-27 Indian Institute Of Technology Delhi Heat exchanger
US7337835B2 (en) 2005-01-25 2008-03-04 Indian Institute Of Technology Delhi Baffle and tube for a heat exchanger
US8067201B2 (en) 2009-04-17 2011-11-29 Bristol-Myers Squibb Company Methods for protein refolding
WO2014155349A2 (fr) * 2013-03-29 2014-10-02 Dr. Reddy's Laboratories Repliement de protéines

Non-Patent Citations (13)

* Cited by examiner, † Cited by third party
Title
BADE, P. D.; KOTU, S. P.; RATHORE, A. S: "Optimization of a refolding step for a therapeutic fusion protein in the quality by design (QbD) paradigm", JOURNAL OF SEPARATION SCIENCE, vol. 35, no. 22, 2012, pages 3160 - 3169
HAMMERSCHMIDT, N.; INTERSTEINER, B.; LINGG, N.; JUNGBAUER, A.: "Continuous precipitation of IgG from CHO cell culture supernatant in a tubular reactor", BIOTECHNOLOGY JOURNAL, 2015
HAMMERSCHMIDT, N.; TSCHELIESSNIG, A.; SOMMER, R.; HELK, B.; JUNGBAUER, A: "Economics of recombinant antibody production processes at various scales: IndustryDstandard compared to continuous precipitation", BIOTECHNOLOGY JOURNAL, vol. 9, no. 6, 2014, pages 766 - 775
MANNALL, G. J.; TITCHENERDHOOKER, N. J.; CHASE, H. A.; DALBY, . A.: "A critical assessment of the impact of mixing on dilution refolding", BIOTECHNOLOGY AND BIOENGINEERING, vol. 93, no. 5, 2006, pages 955 - 963
MONISHA MRIDHA MANDAL ET AL: "Numerical modeling of polystyrene synthesis in coiled flow inverter", MICROFLUIDICS AND NANOFLUIDICS, SPRINGER, BERLIN, DE, vol. 10, no. 2, 3 August 2010 (2010-08-03), pages 415 - 423, XP019876804, ISSN: 1613-4990, DOI: 10.1007/S10404-010-0679-Z *
PAN, S.; ZELGER, M.; HAHN, R.; JUNGBAUER, A: "Continuous protein refolding in a tubular reactor", CHEMICAL ENGINEERING SCIENCE, vol. 116, 2014, pages 763 - 772
PAN, S.; ZELGER, M.; JUNGBAUER, A.; HAHN, R.: "Integrated continuous dissolution, refolding and tag removal of fusion proteins from inclusion bodies in a tubular reactor", JOURNAL OF BIOTECHNOLOGY, vol. 185, no. XX, 2014, pages 39 - 50
RAPHAEL, M.; ROHANI, S.: "Isoelectric precipitation of sunflower protein in an MSMPR precipitator: Modelling of PSD with aggregation", CHEMICAL ENGINEERING SCIENCE, vol. 51, no. 19, 1996, pages 4379 - 4384
RAPHAEL, M.; ROHANI, S.: "Sunflower protein precipitation in a tubular precipitator", THE CANADIAN JOURNAL OF CHEMICAL ENGINEERING, vol. 77, no. 3, 1999, pages 540 - 554
SAXENA, A. K.; NIGAM, K. D. P.: "Coiled configuration for flow inversion and its effect on residence time distribution", AICHE JOURNAL, vol. 30, no. 3, 1984, pages 363 - 368
SCHLEGL, R.; TSCHELIESSNIG, A.; NECINA, R.; WANDL, R.; JUNGBAUER, A.: "Refolding of proteins in a CSTR", CHEMICAL ENGINEERING SCIENCE, vol. 60, no. 21, 2005, pages 5770 - 5780
TAVARE, N. S.; PATWARDHAN, V.: "Agglomeration in a continuous MSMPR crystallizer", AICHE JOURNAL, vol. 38, no. 3, 1992, pages 377 - 384
WARIKOO, V.; GODAWAT, R.: "A new use for existing technology-continuous precipitation for purification of recombination proteins", BIOTECHNOLOGY JOURNAL, 2015

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107759659A (zh) * 2016-08-19 2018-03-06 昂德生物药业有限公司 注射用重组人组织型纤溶酶原激酶衍生物制备用的复性稀释分散装置及其应用
WO2018138566A1 (fr) * 2017-01-27 2018-08-02 Indian Institute Of Technology, Delhi Onduleurs à flux spiralé compacts utiles en tant que mélangeurs en ligne
WO2020095894A1 (fr) * 2018-11-05 2020-05-14 味の素株式会社 Procédé de production d'une protéine repliée à l'aide d'un microréacteur à flux, et appareil de repliement de protéine
US11520310B2 (en) 2019-06-18 2022-12-06 International Business Machines Corporation Generating control settings for a chemical reactor
US11675334B2 (en) 2019-06-18 2023-06-13 International Business Machines Corporation Controlling a chemical reactor for the production of polymer compounds
WO2021111470A1 (fr) * 2019-12-03 2021-06-10 Indian Institute Of Technology Delhi Procédé de préparation protéines thérapeutiques pégylées
CN115003635A (zh) * 2020-02-03 2022-09-02 默克专利股份有限公司 模块化培养室和病毒灭活方法
WO2021230956A1 (fr) * 2020-05-11 2021-11-18 Purdue Research Foundation Synthèse à l'échelle de lomustine dans des conditions d'écoulement continu
EP3984627A1 (fr) 2020-10-19 2022-04-20 Novasep Equipment Solutions Appareil et procédé de purification de biomolécules
WO2022083901A1 (fr) 2020-10-19 2022-04-28 Novasep Equipment Solutions Appareillage et procédé pour la purification de biomolécules

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