Ultra Scale-down Research Articles

Ultra-Scale Down Models: A Review

Ultra scale-down (USD) models have emerged as indispensable tools, addressing challenges in process development and optimization. This review delves into the transformative potential of Ultra Scale-Down (USD) models, exploring their applications, benefits, limitations, and future directions. As industries deal with more complicated manufacturing processes, the integration of USD models into development pipelines becomes crucial. This approach holds the potential to reduce timeframes, cut costs, and enhance the overall efficiency of various processes, contributing to advancements across diverse fields. Envision transforming intricate manufacturing not through colossal machinery but by harnessing the capabilities of tiny vials and microfluidic chips. This encapsulates the essence of ultra scale-down (USD), an innovative approach that downsizes the landscape of process development. Essentially, USD duplicates pivotal elements of large-scale processes on a considerably smaller scale, typically employing milliliter-sized volumes. While USD is still advancing, and its potential to transform process development is evident. Embracing miniaturization, researchers and engineers unleash efficiency, cut costs, and hasten the launch of innovative products. Keywords: Ultra Scale-Down (USD) models, Downstream processing, Cell-based therapies, Early-stage optimization, Real-time applications, Personalized medicine

Integrated ultra scale-down and multivariate analysis of flocculation and centrifugation for enhanced primary recovery

A linked ultra scale-down (USD) flocculation and centrifugation system facilitates rapid bioprocess development and evaluation. These techniques allow studies to be undertaken at high throughput using as little as 50 mL of feed material. They enable the investigation of multiple conditions, thus generating a relatively large amount of data.This study establishes USD flocculation and centrifugation and joins a sequential multivariate data analysis (MVDA) to evaluate the multiparameter effects on primary recovery performance. MVDA techniques such as Principal Component Analysis (PCA) and Partial Least Squares (PLS) were used to handle the complex data sets and investigate the relationships between process parameters and responses. A strategy to assess floc characteristics using PCA was proposed to eliminate the visual inspection, and aid in the analysis, of particle size distribution (PSD) datasets. The PSD of non-sheared and sheared flocs provide good indicators of floc centrifuge performance.The findings show that this USD system can be used to forecast pilot scale performance. The sequential analysis demonstrated that the produced flocs are shear-sensitive in which feed preparation and process shear significantly impact the centrifugation of those flocs. Strong flocs may not necessarily result from high Camp number values (≥ 105) where optimisation of the flocculation chemical parameters is required.The novel integration of the USD systems with MVDA is a powerful platform to optimise and expand process know-how.

Reactor design for continuous monoclonal antibody precipitation based upon micro‐mixing

AbstractBACKGROUNDPrecipitation has been applied for the processing of important therapeutics, including monoclonal antibodies (mAbs). The scale‐up has proven to be a challenging task due to the complexity of the reactions and transport processes involved. This requires a good understanding of the molecular processes underpinning precipitate formation. The aim of this study was to build a micro‐mixing model for the precipitation of a mAb in continuous tubular reactors using ammonium sulphate. The effect of micro‐mixing on precipitate formation (with respect to size, strength, and nature) was evaluated. An ultra scale‐down (USD) centrifugation methodology was applied to determine the ease of precipitate clarification.RESULTSThe results demonstrated that the final mean particle size decreased with increased micro‐mixing, and was obtained with short residence times. Antibody yields in the tubular reactors were consistently above 90% and were shown to be independent of the mixing. Similar particle sizes between a lab and pilot‐scale reactor were correlated with the average energy dissipation rate. The smaller particles obtained from improved micro‐mixing had higher fractal dimensions that correlated with minimal breakage upon exposure to turbulent shear. Precipitates were easily clarified at the USD scale (> 95% clarification), but less so at pilot‐scale (< 80% clarification).CONCLUSIONPrecipitation is a rapid process where the final precipitate properties are controlled by the flow conditions. Therefore, the process can be manipulated to acquire a certain particle size range. A high‐throughput precipitation process is also possible. However, further investigation into large‐scale precipitate recovery is required. © 2020 Society of Chemical Industry

The Use of a Shear Device to Monitor the Stability of a Single-Chain Variable Fragment (scFv) Fusion Protein MFECP1

Antibody based drugs are increasingly being used to treat a vast array of diseases because of their unique affinity to target specific antigen proteins on the surfaces of target cancer cells. Fusions of antibodies and conjugated biopharmaceuticals are progressively being used as this gives the opportunity to target other cytotoxic molecules to unwanted cells. It is critical to ensure these types of drug products are not fragile or uneconomical to produce at a large scale. A very small amount of precious protein solution can be characterised in an Ultra scale-down (USD) shear device to uncover if fusion proteins are prone to shear stress. This article presents how the purified and deglycosylated form of the MFECP1 fusion protein was quantified with an ELISA from 700-50 ng/ml, with a +/- 10% deviation in the standard curve. It also describes how the same MFECP1 fusion protein was analysed to establish the optimum experimental control conditions that were required to observe changes due to hydrodynamic-associated degradation in a shear device. Lastly, it looks at how a first order kinetic relationship can be used to model the rate of MFECP1 fusion protein degradation and how this was used to quantify the rate of protein loss during different shear environments with and without air/liquid interfaces.

Downstream Processing of Chlamydomonas reinhardtii TN72 for Recombinant Protein Recovery.

The green microalga Chlamydomonas reinhardtii is under development as a production host for recombinant proteins and whole-cell therapeutics. In particular, the cell wall-reduced strain TN72 is used as a model organism for protein expression and algal synthetic biology. However, the bioprocessing characteristics of TN72 and other C. reinhardtii strains have yet to be examined. Here we use a TN72 strain expressing a protein-based antibiotic (Pal) to study the scale-up of cell harvest and product recovery. Cell harvest was examined with 100L cultures in two intermittent-discharge continuous-flow disc-stack centrifuges at flow rates of 150–250 L.h−1, as well as with an ultra scale-down (USD) mimic of the centrifuges. Solids recovery exceeded 99.5% and the loss of product to the supernatant was below 2–3%. TN72 is intact following the high shear conditions of the feed zone, however discharge from both disc-stack centrifuges resulted in full cell breakage and in the case of Pal, partial degradation in the subsequent hours. We demonstrated that shake flask cultivation and the USD centrifuge technique can be used to predict the pilot-scale clarification efficiency and product release at the centrifuge inlet for TN72, but not the cell breakage on discharge. This study outlines a number of challenges for scale-up of recombinant protein production in the microalgal host in particular for whole cell therapeutics, but also opportunities for the bioprocessing of intracellular products from TN72.

The prediction of the operating conditions on the permeate flux and on protein aggregation during membrane processing of monoclonal antibodies

The lack of available material during early stage bioprocess development poses numerous processing challenges such as limiting the number of full-scale experiments. Extended fundamental process understanding could be gained with the use of an ultra scale-down (USD) device using as little as 1.7 mL per experimental run. The USD system is used to predict diafiltration and ultrafiltration/diafiltration (UF/DF) performance of a pilot-scale tangential flow filtration (TFF) system, fitted with a flat-sheet cassette, operating at 500-fold larger scale. Both systems were designed by maintaining a volumetric loading of 8.1 L of feed per m2. Permeate flux was predicted for monoclonal antibody solutions with the USD system across a range of transmembrane pressure drops, feed concentrations and flow conditions during diafiltration, and desired retentate concentrations during UF/DF operations. The resulting USD data were in good agreement with the pilot-scale TFF when scaled based on similar shear rates over the membrane surface. Little change in soluble aggregates was observed in both systems but there were significantly higher increases in product turbidity in the USD system. A correlation was established to relate turbidity increase based on the volume fraction of high shear stress zone for USD systems and various pilot-scale TFF systems. The correlation was extended to encompass the processing time and concentration for a wide range of membrane processing challenges in both scales.

Improving the Bioprocessing of ADEPT Fusion Proteins using Ultra Scale-Down Techniques

Improving the Bioprocessing of ADEPT Fusion Proteins using Ultra Scale-Down Techniques

An ultra scale-down method to investigate monoclonal antibody processing during tangential flow filtration using ultrafiltration membranes.

The availability of material for experimental studies is a key constraint in the development of full‐scale bioprocesses. This is especially true for the later stages in a bioprocess sequence such as purification and formulation, where the product is at a relatively high concentration and traditional scale‐down models can require significant volumes. Using a combination of critical flow regime analysis, bioprocess modelling, and experimentation, ultra scale‐down (USD) methods can yield bioprocess information using only millilitre quantities before embarking on highly demanding full‐scale studies. In this study the performance of a pilot‐scale tangential flow filtration (TFF) system based on a membrane flat‐sheet cassette using pumped flow was predicted by devising an USD device comprising a stirred cell using a rotating disc. The USD device operates with just 2.1 cm2 of membrane area and, for example, just 1.7 mL of feed for diafiltration studies. The novel features of the design involve optimisation of the disc location and the membrane configuration to yield an approximately uniform shear rate. This is characterised using computational fluid dynamics for a defined layer above the membrane surface. A pilot‐scale TFF device operating at ~500‐fold larger feed volume and membrane area was characterised in terms of the shear rate derived from flow rate‐pressure drop relationships for the cassette. Good agreement was achieved between the USD and TFF devices for the flux and resistance values at equivalent average shear rates for a monoclonal antibody diafiltration stage.

An ultra scale-down methodology to characterize aspects of the response of human cells to processing by membrane separation operations.

ABSTRACTTools that allow cost‐effective screening of the susceptibility of cell lines to operating conditions which may apply during full scale processing are central to the rapid development of robust processes for cell‐based therapies. In this paper, an ultra scale‐down (USD) device has been developed for the characterization of the response of a human cell line to membrane‐based processing, using just a small quantity of cells that is often all that is available at the early discovery stage. The cell line used to develop the measurements was a clinically relevant human fibroblast cell line. The impact was evaluated by cell damage on completion of membrane processing as assessed by trypan blue exclusion and release of intracellular lactate dehydrogenase (LDH). Similar insight was gained from both methods and this allowed the extension of the use of the LDH measurements to examine cell damage as it occurs during processing by a combination of LDH appearance in the permeate and mass balancing of the overall operation. Transmission of LDH was investigated with time of operation and for the two disc speeds investigated (6,000 and 10,000 rpm or ϵ max ≈ 1.9 and 13.5 W mL−1, respectively). As expected, increased energy dissipation rate led to increased transmission as well as significant increases in rate and extent of cell damage. The method developed can be used to test the impact of varying operating conditions and cell lines on cell damage and morphological changes. Biotechnol. Bioeng. 2017;114: 1241–1251. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

Integrated economic and experimental framework for screening of primary recovery technologies for high cell density CHO cultures.

Increases in mammalian cell culture titres and densities have placed significant demands on primary recovery operation performance. This article presents a methodology which aims to screen rapidly and evaluate primary recovery technologies for their scope for technically feasible and cost‐effective operation in the context of high cell density mammalian cell cultures. It was applied to assess the performance of current (centrifugation and depth filtration options) and alternative (tangential flow filtration (TFF)) primary recovery strategies. Cell culture test materials (CCTM) were generated to simulate the most demanding cell culture conditions selected as a screening challenge for the technologies. The performance of these technology options was assessed using lab scale and ultra scale‐down (USD) mimics requiring 25–110mL volumes for centrifugation and depth filtration and TFF screening experiments respectively. A centrifugation and depth filtration combination as well as both of the alternative technologies met the performance selection criteria. A detailed process economics evaluation was carried out at three scales of manufacturing (2,000L, 10,000L, 20,000L), where alternative primary recovery options were shown to potentially provide a more cost‐effective primary recovery process in the future. This assessment process and the study results can aid technology selection to identify the most effective option for a specific scenario.

Evaluation of options for harvest of a recombinant E. Coli fermentation producing a domain antibody using ultra scale‐down techniques and pilot‐scale verification

Ultra scale‐down (USD) methods operating at the millilitre scale were used to characterise full‐scale processing of E. coli fermentation broths autolysed to different extents for release of a domain antibody. The focus was on the primary clarification stages involving continuous centrifugation followed by depth filtration. The performance of this sequence was predicted by USD studies to decrease significantly with increased extents of cell lysis. The use of polyethyleneimine reagent was studied to treat the lysed cell broth by precipitation of soluble contaminants such as DNA and flocculation of cell debris material. The USD studies were used to predict the impact of this treatment on the performance and here it was found that the fermentation could be run to maximum productivity using an acceptable clarification process (e.g., a centrifugation stage operating at 0.11 L/m2 equivalent gravity settling area per hour followed by a resultant required depth filter area of 0.07 m2/L supernatant). A range of USD predictions was verified at the pilot scale for centrifugation followed by depth filtration. © 2016 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 32:382–392, 2016

Creation of an ultra scale-down bioreactor mimic for rapid development of lignocellulosic enzymatic hydrolysis processes.

BACKGROUNDCellulosic bioethanol processes involve several steps, all of which require experimental optimisation. A significant aid to this research would be a validated ultra scale‐down (USD) model that could be used to perform rapid, wide ranging screening and optimisation experiments using limited materials under process relevant conditions.RESULTSIn this work, the use of 30 mL shaken conical tubes as a USD model for an enzymatic hydrolysis process is established. The approach is demonstrated for the hydrolysis of distillers' dried grains with solubles (DDGS). Results from the USD tubes closely mimic those obtained from 4 L stirred tanks, in terms of the rate, composition and concentrations of sugars released, representing an 80‐fold scale reduction. The utility of the USD approach is illustrated by investigating factors that may be limiting hydrolysis yields at high solids loadings. Washing the residual solids periodically during hydrolysis allowed 100% of the available sugar to be hydrolysed using commercially available enzymes.CONCLUSIONThe results demonstrate that the USD system reported successfully mimics the performance of conventional stirred tanks under industrially relevant conditions. The utility of the system was confirmed through its use to investigate performance limitation using a commercially relevant feedstock. © 2015 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A modular approach for the ultra-scale-down of depth filtration

Depth filtration is employed in the production of many biological products. Although high throughput process development is available for most purification operations, options for high throughput depth filtration are limited. In this report, we describe the development and qualification of an ultra scale-down (USD) device for high throughput depth filtration. The system enables the parallel evaluation of eight single- or multi-layer depth filters approximately 0.2cm2 in cross-sectional area, greater than two logs smaller than the smallest commercially available devices. To assess precision, scalability, modularity, and media heterogeneity in USD depth filtration, several types of adsorbent media were qualified in the USD device, with filtration performance compared to lab-scale devices (cross-sectional area: 23–25cm2). The performance was further investigated with several high throughput clarity measurements, including particle characterization of filtrate fractions. Historically, depth filtration development has been limited by the amount of material required to run lab-scale filters. The described high throughput technology enables the screening of filter media early in development, optimization of operating parameters, and coupling of upstream operations to filtration performance. These high throughput capabilities will create efficiencies for development and manufacturing that were unattainable previously and create an important tool for screening novel biological entities.

Scale-down characterization of post-centrifuge flocculation processes for high-throughput process development.

AbctractFlocculation unit operations are being revisited as a strategy to ease the burden posed on clarification and purification operations by the increasingly high cell density cultures used in the biopharmaceutical industry. The purpose of this study was to determine the key process parameters impacting flocculation scale-up and use this understanding to develop an automated ultra-scale down (USD) method for the rapid characterization of flocculation at the microliter scale. The conditions under which flocculation performance of a non-geometrically similar vessel three orders of magnitude larger can be mimicked by the USD platform are reported. Saccharomyces cerevisiae clarified homogenate was flocculated with poly(ethyleneimine) (PEI) to remove the residual solids remaining in the centrate. Flocculant addition time modulated flocculation performance depending on the predominant mixing time scale (i.e. macro-, meso- or micromixing). Particle growth and breakage was mimicked at the two flocculation scales by the average turbulent energy dissipation (εavg) and impeller tip speed (vtip) scale-up bases. The results obtained were used to develop an USD method. The USD method proposed uses constant εavg as the scale-up basis under a micromixing controlled regime. These conditions mimicked the STR flocculation performance within a ±5% error margin. Operation in the mesomixing regime led to particle size deviations between the flocculation scales of ≤50 %. These results, in addition to the microscopic observations made, demonstrate the USD system presented in this work can produce process-relevant flocculated material at the microliter scale under the correct operating conditions.

An ultra scale‐down approach to study the interaction of fermentation, homogenization, and centrifugation for antibody fragment recovery from rec E. coli

Escherichia coli is frequently used as a microbial host to express recombinant proteins but it lacks the ability to secrete proteins into medium. One option for protein release is to use high-pressure homogenization followed by a centrifugation step to remove cell debris. While this does not give selective release of proteins in the periplasmic space, it does provide a robust process. An ultra scale-down (USD) approach based on focused acoustics is described to study rec E. coli cell disruption by high-pressure homogenization for recovery of an antibody fragment (Fab') and the impact of fermentation harvest time. This approach is followed by microwell-based USD centrifugation to study the removal of the resultant cell debris. Successful verification of this USD approach is achieved using pilot scale high-pressure homogenization and pilot scale, continuous flow, disc stack centrifugation comparing performance parameters such as the fraction of Fab' release, cell debris size distribution and the carryover of cell debris fine particles in the supernatant. The integration of fermentation and primary recovery stages is examined using USD monitoring of different phases of cell growth. Increasing susceptibility of the cells to disruption is observed with time following induction. For a given recovery process this results in a higher fraction of product release and a greater proportion of fine cell debris particles that are difficult to remove by centrifugation. Such observations are confirmed at pilot scale.

An ultra scale‐down characterization of low shear stress primary recovery stages to enhance selectivity of fusion protein recovery from its molecular variants

Fusion proteins offer the prospect of new therapeutic products with multiple functions. The primary recovery is investigated of a fusion protein consisting of modified E2 protein from hepatitis C virus fused to human IgG1 Fc and expressed in a Chinese hamster ovary (CHO) cell line. Fusion protein products inevitably pose increased challenge in preparation and purification. Of particular concerns are: (i) the impact of shear stress on product integrity and (ii) the presence of product-related contaminants which could prove challenging to remove during the high resolution purification steps. This paper addresses the use of microwell-based ultra scale-down (USD) methods to develop a bioprocess strategy focused on the integration of cell culture and cell removal operations and where the focus is on the use of operations which impart low shear stress levels even when applied at eventual manufacturing scale. An USD shear device was used to demonstrate that cells exposed to high process stresses such as those that occur in the feed zone of a continuous non-hermetic centrifuge resulted in the reduction of the fusion protein and also the release of glycosylated intracellular variants. In addition, extended cell culture resulted in release of such variants. USD mimics of low shear stress, hydrohermetic feed zone centrifugation and of depth filtration were used to demonstrate little to no release during recovery of these variants with both results verified at pilot scale. Furthermore, the USD studies were used to predict removal of contaminants such as lipids, nucleic acids, and cell debris with, for example, depth filtration delivering greater removal than for centrifugation but a small (~10%) decrease in yield of the fusion protein. These USD observations of product recovery and carryover of contaminants were also confirmed at pilot scale as was also the capacity or throughput achievable for continuous centrifugation or for depth filtration. The advantages are discussed of operating a lower yield cell culture and a low shear stress recovery process in return for a considerably less challenging purification demand.

An ultra scale‐down approach to assess the impact of the choice of recombinant P. pastoris strain on dewatering performance in centrifuges

Pichia pastoris is becoming a desirable host in the biopharmaceutical industry for therapeutics production. It grows on methanol to high cell densities ≥100 g DCW/L and secretes foreign proteins at high titers. However, the culture conditions to reach high cell densities pose a challenge to the processability by primary recovery operations, in particular centrifugation, used for cell removal. This work aims to assess the impact of recombinant P. pastoris strain selection on centrifugal dewatering. Normally, the choice of P. pastoris recombinant strain is based on best target protein expression levels; however, it is unknown whether the choice of strain will have an impact on performance of centrifugation operation. To achieve this aim, a previously developed laboratory ultra-scale down (USD) methodology that successfully predicted centrifugal dewatering of pilot-scale disk-type machines, was used in this work. Two recombinant P. pastoris strains, namely a X-33 and a glycoengineered Pichia strain, were used to perform fermentations secreting different products. The resulting harvested fermentation culture properties were analyzed and the dewatering performances of a pilot- and a large-scale disk-type centrifuge were evaluated using the USD methodology. The choice of P. pastoris strain was found to have a considerable impact on dewatering performance, with P. pastoris X-33 strain reaching better dewatering levels than the glycoengineered strain. The USD method proved to be a useful tool to determine optimal conditions under which the large scale centrifuge needed to be operated, reducing the need for repeated pilot-scale runs during early stages of process development for therapeutic products.

Use of focused acoustics for cell disruption to provide ultra scale‐down insights of microbial homogenization and its bioprocess impact—recovery of antibody fragments from rec E. coli

An ultra scale-down (USD) device that provides insight of how industrial homogenization impacts bioprocess performance is desirable in the biopharmaceutical industry, especially at the early stage of process development where only a small quantity of material is available. In this work, we assess the effectiveness of focused acoustics as the basis of an USD cell disruption method to mimic and study high-pressure, step-wise homogenization of rec Escherichia coli cells for the recovery of an intracellular protein, antibody fragment (Fab'). The release of both Fab' and of overall protein follows first-order reaction kinetics with respect to time of exposure to focused acoustics. The rate constant is directly proportional to applied electrical power input per unit volume. For nearly total protein or Fab' release (>99%), the key physical properties of the disruptate produced by focused acoustics, such as cell debris particle size distribution and apparent viscosity show good agreement with those for homogenates produced by high-pressure homogenization operated to give the same fractional release. The only key difference is observed for partial disruption of cells where focused acoustics yields a disruptate of lower viscosity than homogenization, evidently due to a greater extent of polynucleic acids degradation. Verification of this USD approach to cell disruption by high-pressure homogenization is achieved using USD centrifugation to demonstrate the same sedimentation characteristics of disruptates prepared using both the scaled-down focused acoustic and the pilot-scale homogenization methods for the same fraction of protein release.

Prediction and verification of centrifugal dewatering of P. pastoris fermentation cultures using an ultra scale‐down approach

Recent years have seen a dramatic rise in fermentation broth cell densities and a shift to extracellular product expression in microbial cells. As a result, dewatering characteristics during cell separation is of importance, as any liquor trapped in the sediment results in loss of product, and thus a decrease in product recovery. In this study, an ultra scale-down (USD) approach was developed to enable the rapid assessment of dewatering performance of pilot-scale centrifuges with intermittent solids discharge. The results were then verified at scale for two types of pilot-scale centrifuges: a tubular bowl equipment and a disk-stack centrifuge. Initial experiments showed that employing a laboratory-scale centrifugal mimic based on using a comparable feed concentration to that of the pilot-scale centrifuge, does not successfully predict the dewatering performance at scale (P-value <0.05). However, successful prediction of dewatering levels was achieved using the USD method (P-value ≥0.05), based on using a feed concentration at small-scale that mimicked the same height of solids as that in the pilot-scale centrifuge. Initial experiments used Baker's yeast feed suspensions followed by fresh Pichia pastoris fermentation cultures. This work presents a simple and novel USD approach to predict dewatering levels in two types of pilot-scale centrifuges using small quantities of feedstock (<50 mL). It is a useful tool to determine optimal conditions under which the pilot-scale centrifuge needs to be operated, reducing the need for repeated pilot-scale runs during early stages of process development.

Characterization and feasibility of a miniaturized stirred tank bioreactor to perform E. coli high cell density fed‐batch fermentations

The use of small scale bioreactors that are mechanically and functionally similar to large scale reactors is highly desirable to accelerate bioprocess development because they enable well-defined scale translations. In this study, a 25-mL miniaturized stirred tank bioreactor (MSBR) has been characterized in terms of its power input, hydrodynamics, and volumetric oxygen transfer coefficient (k(L)a) to assess its potential to grow high cell density (HCD) cultures using adequate scale-down criteria. Engineering characterization results show scale down, based on matched specific power input (P(G)/V), is feasible from a 20-L pilot scale stirred tank bioreactor. Results from fed-batch fermentations performed using Fab' producing E. coli W3110 at matched (P(G)/V) in the MSBR and 20-L STR demonstrated that the MSBR can accurately scale down the 20-L fermentation performance in terms of growth and Fab' production. Successful implementation of a fed-batch strategy in the MSBR resulted in maximum optical density of ca. 114 and total Fab' concentration of 940 μg/mL compared with ca. 118 and 990 μg/mL in 20-L STR. Furthermore, the use of the MSBR in conjunction with primary recovery scale-down tools to assess the harvest material of both reactors showed comparable shear sensitivity and centrifugation performance. The conjoint use of the MSBR with ultra scale-down (USD) centrifugation mimics can provide a cost-efficient manner in which to design and develop bioprocesses that account for good upstream performance as well as their manufacturability downstream.