Adapting viral safety assurance strategies to continuous processing of biological products

Johnson, Sarah; Brown, Matthew R.; Lute, Scott; Brorson, Kurt

doi:10.1002/bit.26245

Cited by 21 publications

(30 citation statements)

References 54 publications

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“…Non-enveloped viruses are generally smaller and more stable than enveloped viruses. Considering the variation in structure and stability, non-enveloped viruses are conventionally removed through filtration or chromatography, [3][4][5][6][7][8][9][10][11] while enveloped viruses are inactivated through chemical or physical processes 4,[12][13][14] in therapeutic protein manufacturing.…”

Section: Virus Biochemistrymentioning

confidence: 99%

Arginine‐enveloped virus inactivation and potential mechanisms

Meingast

Heldt

2019

Biotechnology Progress

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Arginine synergistically inactivates enveloped viruses at a pH or temperature that does little harm to proteins, making it a desired process for therapeutic protein manufacturing. However, the mechanisms and optimal conditions for inactivation are not fully understood, and therefore, arginine viral inactivation is not used industrially. Optimal solution conditions for arginine viral inactivation found in the literature are high arginine concentrations (0.7–1 M), a time of 60 min, and a synergistic factor of high temperature (≥40°C), low pH (≤pH 4), or Tris buffer (5 mM). However, at optimal conditions full inactivation does not occur over all enveloped viruses. Enveloped viruses that are resistant to arginine often have increased protein stability or membrane stabilizing matrix proteins. Since arginine can interact with both proteins and lipids, interaction with either entity may be key to understanding the inactivation mechanism. Here, we propose three hypotheses for the mechanisms of arginine induced inactivation. Hypothesis 1 describes arginine‐induced viral inactivation through inhibition of vital protein function. Hypothesis 2 describes how arginine destabilizes the viral membrane. Hypothesis 3 describes arginine forming pores in the virus membrane, accompanied by further viral damage from the synergistic factor. Once the mechanisms of arginine viral inactivation are understood, further enhancement by the addition of functional groups, charges, or additives may allow the inactivation of all enveloped viruses in mild conditions.

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Section: Virus Biochemistrymentioning

confidence: 99%

Arginine‐enveloped virus inactivation and potential mechanisms

Meingast

Heldt

2019

Biotechnology Progress

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“…CFD may provide a useful predictor of scale-up or scaledown performance. In particular, it may be to change throughput by changing the flow path inner diameter while 25 A small-scale JIB would also fit better into a smallscale integrated process. If it can be established on a demonstration scale that a continuous viral inactivation unit process is just as robust as fed batch step, with lower overall cost of goods and better ease of use, then developing larger-scale (500 L to 1000 L) models may be desirable.…”

Section: Scale Down Model Using Cfdmentioning

confidence: 99%

“…Designers should also evaluate reactor materials to characterize variations (possibly very small) of temperature and pH within the reactor chamber, as suggested by regulatory agencies. 25 In a design that is robust and highly reproducible, conducting these studies should not be unduly difficult. The 3D printing technology is evolving rapidly.…”

Section: Other Challengesmentioning

confidence: 99%

Design, construction, and optimization of a novel, modular, and scalable incubation chamber for continuous viral inactivation

Orozco

Godfrey

Coffman

et al. 2017

Biotechnology Progress

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We designed, built or 3D printed, and screened tubular reactors that minimize axial dispersion to serve as incubation chambers for continuous virus inactivation of biological products. Empirical residence time distribution data were used to derive each tubular design's volume equivalent to a theoretical plate (VETP) values at a various process flow rates. One design, the Jig in a Box (JIB), yielded the lowest VETP, indicating optimal radial mixing and minimal axial dispersion. A minimum residence time (MRT) approach was employed, where the MRT is the minimum time the product spends in the tubular reactor. This incubation time is typically 60 minutes in a batch process. We provide recommendations for combinations of flow rates and device dimensions for operation of the JIB connected in series that will meet a 60-min MRT. The results show that under a wide range of flow rates and corresponding volumes, it takes 75 ± 3 min for 99% of the product to exit the reactor while meeting the 60-min MRT criterion and fulfilling the constraint of keeping a differential pressure drop under 5 psi. Under these conditions, the VETP increases slightly from 3 to 5 mL though the number of theoretical plates stays constant at about 1326 ± 88. We also demonstrated that the final design volume was only 6% ± 1% larger than the ideal plug flow volume. Using such a device would enable continuous viral inactivation in a truly continuous process or in the effluent of a batch chromatography column. Viral inactivation studies would be required to validate such a design. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:954-965, 2017.

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“…Furthermore, it is one of the most important viral clearance steps addressing the enveloped murine leukemia virus, the relevant model virus for retrovirus-like particles found in the production cell lines like Chinese hamster ovary (CHO) cells (CMC Biotech Working Group, 2009). So far, the viral clearance validation of continuously operated downstream process steps has been discussed (Johnson, Brown, Lute, & Brorson, 2017) but not investigated in detail. Already, existing batch techniques have to be adapted to the continuous environment.…”

Section: Introductionmentioning

confidence: 99%

Virus study for continuous low pH viral inactivation inside a coiled flow inverter

David

Maiser

Lobedann

et al. 2018

Biotech & Bioengineering

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Continuous processing for the production of monoclonal antibodies (mAb) gains more and more importance. Several solutions exist for all the necessary production steps, leading to the possibility to build fully continuous processes. Low pH viral inactivation is a part of the standard platform process for mAb production. Consequently, Klutz et al. introduced the coiled flow inverter (CFI) as a tool for continuous low pH viral inactivation. Besides theoretical calculations of viral reduction, no viral clearance study has been presented so far. In addition, the validation of continuous viral clearance is often neglected in the already existing studies for continuous processing. This study shows in detail the development and execution of a virus study for continuous low pH viral inactivation inside a CFI. The concept presented is also valid for adaptation to other continuous viral clearance steps. The development of this concept includes the technical rationale for an experimental setup, a valid spiking procedure, and finally a sampling method. The experimental results shown represent a viral study using xenotropic murine leukemia virus as a model virus. Two different protein A (ProtA) chromatography setups with varying pH levels were tested. In addition, one of these setups was tested against a batch experiment utilizing the same process material. The results show that sufficient low pH viral inactivation (decadic logarithm reduction value >4) was achieved in all experiments. Complete viral inactivation took place within the first 14.5 min for both continuous studies and the batch study, hence showing similar results. This study therefore represents a successful virus study concept and experiment for a continuous viral inactivation step. Moreover, it was shown that the transfer from batch results to the continuous process is possible. This is accomplished by the narrow residence time distribution of the CFI, showing how close the setup approaches the ideal plug flow and with that batch operation. Biotechnology and Bioengineering. 2019;116:857-869. wileyonlinelibrary.com/journal/bit Symbols: d, decadic logarithm of the serial dilution steps; D, sample predilution; n, total number of analyzed cell culture wells; n p , number of virus-positive wells; p, probability value; P i , summation of virus-positive cell cultures within the virus transition area; R w , relative width; v, decadic logarithm of the volume conversion factor; V, overall volume of the virus-containing sample; V w , analyzed sample volume; Y 0 , decadic logarithm of the highest dilution of the virus-containing sample; Θ 0.005 , dimensionless time point where 0.5% of the maximum dimensionless concentration is reached; Θ 0.995 , dimensionless time point where 99.5% of the maximum dimensionless concentration is reached. K E Y W O R D S coiled flow inverter, continuous processing, low pH, monoclonal antibodies, viral inactivation

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Adapting viral safety assurance strategies to continuous processing of biological products

Cited by 21 publications

References 54 publications

Arginine‐enveloped virus inactivation and potential mechanisms

Arginine‐enveloped virus inactivation and potential mechanisms

Design, construction, and optimization of a novel, modular, and scalable incubation chamber for continuous viral inactivation

Virus study for continuous low pH viral inactivation inside a coiled flow inverter

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