Protein A chromatography is a key unit operation in biopharmaceutical purification, widely used to capture monoclonal antibodies (mAbs) from harvest cell culture fluid (HCCF). In addition to its primary function, protein A chromatography is also frequently tested for its ability to remove viruses. Although protein A specifically binds mAbs and should not interact with viruses, some studies have shown that logarithmic reduction values (LRVs) can vary between 1 and 4 log10 for retroviruses. This variance has been shown to depend on the mAb and feedstock compositions. However, it has not been evaluated so far which feedstock ingredients affect virus removal. To address this knowledge gap, a data set of virus spiking studies was analyzed and revealed a potential correlation between DNA content in HCCF and LRV of protein A chromatography. To evaluate this hypothesis, experiments with murine leukemia virus (MuLV) and retrovirus like particles (RVLP) spiked starting materials of three different mAbs were performed. In general, higher HCCF DNA levels were found to enhance LRV for both virus particles. A trend that was confirmed with RVLP spiked pure mAb. In addition, protein A chromatography washing buffers containing a detergent or a reducing agent were shown to improve LRV in RVLP spiked experiments. This study demonstrates the effect of DNA concentration on the virus removal capabilities of protein A chromatography. Findings that are especially useful when DNA is removed at early stages of the downstream process, for example by using functionalized depth filters or precipitation.
Recombinant adeno-associated virus production for gene therapy applications commonly relies on the Spodoptera frugiperda 9 (Sf9) baculovirus infection platform. The process encompasses cell expansion from flasks through intermediate-scale bioreactors to large-scale production. A critical challenge in this process is suboptimal cell proliferation during the expansion phase, which significantly impacts subsequent virus production. This study evaluates the effects of different single-use bioreactor bags on Sf9 cell growth, focusing on two potential issues: incompatibility between Sf9 cells and bag materials, and the presence of leachables and extractables from the bag. Single-use rocking motion bioreactor bag configuration, including material- and surface-related attributes, was associated with substantial differences in Sf9 cell expansion performance. While growth kinetics in shake flasks and the Pall Allegro XRS system were comparable, reduced growth rates and delayed proliferation were observed in Cytiva Cellbags. The comparable growth observed in shake flasks and the Pall XRS system, together with the lack of recovery after rocking-parameter adjustments in Cellbags, suggests that bag-associated material and surface-related factors, rather than rocking motion alone, contributed substantially to the observed growth differences. Lower bag occupancy further exacerbated growth inhibition. Washing studies showed no improvement in new bags, whereas partial recovery after initial use suggested transient bag conditioning or surface-related effects. Analytical extractables and leachables profiling was not performed; therefore, leachable-mediated contributions cannot be fully excluded. These findings highlight the importance of single-use bag selection and occupancy optimization for robust and scalable Sf9 cell expansion processes.
Low-pH viral inactivation (LPVI) is a widely used, validated unit operation for enveloped virus clearance in antibody manufacturing. However, acid exposure during Protein A (ProA) elution and subsequent low-pH holds can destabilize some antibodies and Fc-containing modalities, increasing aggregation risk and product loss-a challenge that is particularly acute for multi-specific antibody formats and other pH-labile Fc-containing modalities. Mild-pH ProA capture strategies protect such molecules, but the resulting higher-pH eluates often require re-acidification to meet conventional LPVI setpoints, reintroducing low-pH stress. Here we evaluate a dual-mechanism strategy in which mild acidity is combined with low concentrations of non-ionic detergent to accelerate inactivation without prolonged exposure to strongly acidic conditions. Using xenotropic murine leukemia virus (X-MuLV) spiked into ProA eluates, 0.10% N-methylglucamide (Mega-10) at pH 3.91 reduced infectivity to below the assay limit of detection within 2.5 min (log10 reduction factor [LRF] ≥ 5.40), whereas low pH alone or detergent alone produced substantially lower reductions over the same interval. Similarly, 0.15% Mega-10 at pH 4.05 achieved nondetectable infectivity within 10 min (LRF ≥ 5.81). Comparable time compression was observed with Tween 80 or Tween 20 at pH ~3.97, reducing time-to-nondetectable infectivity from 120 min (low pH alone) to 5 min. The operating window was tunable through paired adjustment of detergent concentration and pH, and feasibility was demonstrated at pH 4.56 with low detergent levels, offering a practical solution for pH-labile products and higher-pH eluates.
Continuous chromatography for Protein A capture of monoclonal antibodies (mAbs) and related biotherapeutics is gaining popularity for higher throughput and reduced costs but introduces challenges for development and modeling. We present a streamlined, single-breakthrough-curve approach that predicts maximum and operating binding capacities (MBC and OBC, respectively) for continuous Protein A chromatography of monoclonal antibodies and derivatives. Our empirically-calibrated model-intended for early-stage, material-limited process development-enables accurate OBC prediction (root mean squared error [RMSE]≤ 5% for four distinct proteins) using less than 1 g of protein. This approach reduces the experimental timeline by ~2 weeks and is best suited for rapid screening and initial process optimization, not full design-space determination. Results confirm the model's accuracy across molecules, with clear practical advantages for accelerating preclinical pipeline advancement.
Perfusion bioreactor systems enable intensified bioprocessing with high cell densities and volumetric productivities in monoclonal antibody (mAb) manufacturing. However, their widespread adoption remains constrained by the high consumption of chemically defined cell culture media. In this study, we introduce a multi-stage electrokinetic waste separation process that regenerates spent media, recovering up to 87.5% of the spent-medium volume for reuse. Using a mock-perfusion model, the regeneration-recycle processes were shown to sustain high-density Chinese hamster ovary cell growth, compared with spent media recycling without regeneration. Integration into a self-recycling perfusion bioreactor (350 mL working volume) operating with 75% regenerated media sustained the target viable cell density and antibody titer, with limited reductions to cell-specific growth rate (~31%) and cell-specific productivity (~9%), supporting the feasibility of high-ratio media recycling. Techno-economic modeling of large-scale scenarios revealed that media regeneration could offer a path to $15/g COGs, thereby increasing the accessibility of mAbs. Techno-economic modeling of hypothetical large-scale manufacturing scenarios indicated that the experimentally demonstrated process could reduce cost of goods (COG) by approximately 9%, and identified a theoretical pathway toward ~$15/g COG contingent upon multi-cycle regeneration processes that could be developed in the future. These results support regenerative perfusion as a promising process-intensification strategy that, with further development, may be adaptable to other mammalian cell culture platforms where media cost and sustainability are critical bottlenecks.
Reliable assessment of cellular state remains a challenge in cell therapy manufacturing and regenerative medicine. Proliferative capacity, senescence, and metabolic activity are traditionally measured using labor-intensive assays, limiting timely and integrated assessment of cellular state. Here, we apply attenuated total reflection-Fourier transform infrared spectroscopy coupled with multivariate analysis to assess whether intrinsic vibrational fingerprints of mesenchymal stem cells reflect coordinated functional changes during in vitro expansion. By integrating proliferation kinetics, metabolite fluxes, and senescence-associated phenotype into a composite functional score, we mapped progressive functional decline across passages onto infrared spectral changes. Spectral alterations were associated with shifts in lipid-related and protein-associated regions as senescence accumulated. Despite strong donor-specific differences, we identified conserved spectral fingerprint regions that tracked functional decline across donor samples analyzed. These shared biochemical signatures were associated with the prediction of senescence progression and the estimation of functional passage in an independent donor sample. Together, our results show that intrinsic vibrational fingerprints capture coordinated biochemical changes during cellular decline and provide a label-free, rapid means to assess cellular state.
Abstract Recombinant adeno‐associated virus (rAAV) production commonly yields a high proportion of empty and partially filled capsids, necessitating downstream purification to enrich full capsids. Among purification approaches, anion‐exchange chromatography (AEX) has emerged as a scalable alternative to ultracentrifugation. In this study, a structured and scalable development strategy for AAV5 AEX purification is presented. A high‐throughput, ultra–scale‐down platform using 200 μL RoboColumns™ was established to enable rapid screening of salt systems, additives, and operating conditions with minimal material requirements. Dual‐salt systems were evaluated to decouple ionic strength from ion‐specific effects, leading to the identification of a magnesium‐only system (MgCl 2 /MgSO 4 ) that improved both enrichment and recovery. Optimized conditions achieved >85% full capsids with robust recovery and >90% full capsids in peak fractions. Scalability was demonstrated through strong agreement between ultra scale‐down (200 μL) and scale‐down (1 mL) formats. A QbD‐aligned framework was applied to define the purity–recovery design space and guide selection of operating conditions based on target product profile. The optimized gradient process was translated into a manufacturing‐relevant isocratic step format, with systematic evaluation of buffer conductivity sensitivity and load robustness to address large‐scale operational constraints.
Development of therapeutic bispecific antibodies (BsAbs) poses significant manufacturability challenges associated with correct chain pairing and an increased diversity of impurities. Here we address this with a product quality assessment during cell line development (CLD), forgoing the need for genetic characterization during cell line screening. Using Chinese Hamster Ovary (CHO) cell lines expressing a panel of monoclonal antibodies (mAb) and BsAbs, we demonstrate the highly specific binding of SpotLight™ Huλ, Huĸ and HuFc reagents to their target chain isotypes (lambda or kappa light chain and Fc regions) during single cell cloning workflows on a Bruker Beacon® Optofluidic System. We then demonstrate combinatorial use of SpotLight™ Huλ and Huĸ reagents as an early product quality indicator on a CHO cell line expressing a BsAb with dual kappa-lambda light chain configuration. Cell lines presenting equal binding profiles for each SpotLight assay exhibited platform-typical heterodimer levels with an average of 87.26% following fed-batch bioreactor production assessment. Conversely, cell lines biased towards either SpotLight™ assay resulted in lower than desired heterodimer (0%-84.23%) and increased associated impurities. This study demonstrates the application of dual SpotLight™ assays as an identifier of CHO cell lines expressing a kappa-lambda light chain BsAb with a high heterodimer percentage. This allows cell lines with optimal product quality to be identified earlier in CLD, resulting in fewer cell lines being progressed to resource-intensive fed-batch studies and subsequent product quality assessment. Furthermore, this assay provides an opportunity to characterize impurity expression biases prior to developing downstream purification processes.
The sustainable production of hydroxy fatty acids, such as 10-hydroxystearic acid (10-HSA), by biocatalysis is a promising alternative to petrochemical and castor oil-derived products. However, industrial implementation still requires an efficient and scalable process for biocatalyst production by fermentation, which precedes the biotransformation to 10-HSA. In general, industrial biocatalysis is commonly performed using whole cells, which are cheaper than purified enzymes. Still, the fermentation process itself is a major cost contributor requiring a high-yielding and scalable process. Here, we established and scaled a lactose-induced fed-batch process to produce an Escherichia coli BL21 (DE3) based whole-cell biocatalyst containing an oleate hydratase from Stenotrophomonas nitritireducens. The two-phased process employs an initial growth phase on glucose, followed by an induced feed phase using glycerol and lactose. When comparing two different growth rates during enzyme expression, a higher growth rate was found beneficial, resulting in a higher biomass concentration of 69.2 ± 0.5 g L-1 and a yield increase of 80 % while maintaining biocatalyst activity at >85 % conversion. This highlights the importance of process design variables, such as growth rate settings, for a high-yielding and economic process. Feasibility of the process was demonstrated by scaling into a 150 L bioreactor, achieving a biomass concentration of 60.6 ± 0.4 g L-1 with a yield of 0.44 gC,Biomass gC -1 and 91.5 ± 1.4 % conversion. Supplemented with an initial test of suitable unit operations for technical biomass separation, this work provides a fermentation route for a whole-cell oleate hydratase biocatalyst, paving the way for further scaling toward industrial 10-HSA production.
Physiologically relevant lung-on-chip (LoC) systems remain constrained by polydimethylsiloxane (PDMS) limitations including small-molecule adsorption and mechanical deformation of device microchannels under perfusion, single-channel architectures limiting parallel tissue culture, and absence of organized stromal and immune compartments. Existing platforms predominantly model the distal airway, leaving the upper airway, the primary site of respiratory viral entry, insufficiently addressed as an in vitro tissue model for preclinical drug evaluation. We report a six-well, 3D-printed LoC platform fabricated using a photopolymer resin that addresses these limitations. Computational fluid dynamics (CFD)-guided design ensured uniform perfusion across all six wells, enabling parallel tissue culture within a single device. The photopolymer architecture eliminated mechanical deformation of the device microchannels under prolonged perfusion and demonstrated significantly reduced small-molecule adsorption relative to PDMS, improving quantitative reliability for drug evaluation workflows. Human tracheobronchial epithelial cells differentiated at air-liquid interface expressed MUC5AC and cytokeratin-14, formed continuous ZO-1 tight junctions, and achieved a transepithelial electrical resistance of ~800 Ω·cm2, consistent with physiological barrier integrity. A GelMA-based stromal compartment, fabricated by extrusion bioprinting of lung fibroblast- and THP-1 monocyte-laden bioink, established a layered epithelial-stromal-immune architecture. Functional validation demonstrated IL-6 and IL-8 elevation following SARS-CoV-2 spike S1 stimulation, IFN-γ-driven modulation of apical ACE2 expression, and coordinated TNF-α, IL-6, and IL-8 release during Influenza A infection, all suppressed by oseltamivir treatment. This platform integrates CFD-optimized multi-well device fabrication, extrusion bioprinting of stromal constructs, and photopolymer resin-based microfluidic manufacturing into a reproducible, human-relevant upper airway model for respiratory infection research and preclinical antiviral drug evaluation.
The vaccine and viral vector industry is growing at an accelerated rate. To improve harvest and purification processes, the development of continuous membrane-based operations, such as normal flow filtration (NFF) and single pass tangential flow filtration (SPTFF) for concentration were explored. This work was conducted using two model viruses, non-enveloped porcine parvovirus (PPV) and enveloped Suid herpesvirus (SuHV). The viruses are in the same family as the gene therapy vectors adeno associated virus and herpes simplex virus, respectively. SPTFF design started with batch TFF for membrane selection. Hollow fiber membranes with a 100 and 300 kDa molecular weight cut off were defined for PPV and SuHV SPTFF operations, respectively. The SPTFF runs for PPV did not provide any concentration of the virus and low protein and DNA removal, unlike batch TFF. Two hollow fiber membranes run at 10 mL/min and 2 psi were the best condition for SuHV concentration, with approximately 100-fold titer concentration and protein and DNA removal of 37% ± 5% and 32% ± 8%, respectively. This concentration was superior to the batch TFF and indicated a strong dependence on flow rate and transmembrane pressure. For NFF, filters selection and performance tests were carried out for NFF of PPV and SuHV, as well as cleaning protocols for hollow fiber membranes. The ultimate goal is to integrate this work into the continuous purification of viral vectors produced in mammalian cell cultures to reduce costs and increase throughput.
Maintaining consistent quality in the manufacturing of biotherapeutic proteins in mammalian cell culture is challenging, with unplanned deviations causing inconsistencies and potential batch failure. Current methods for monitoring and controlling critical process parameters (CPPs) rely on slow, labor-intensive offline analyses. This is particularly problematic in upstream manufacturing, where infrequent measurements hinder real-time CPP monitoring and result in suboptimal control. This study developed and implemented a robust, standardized framework that integrates Raman spectroscopy with real-time machine learning models and bioreactor control via OPC-UA, enabling seamless communication and effective control of mammalian cell culture CPPs. High-accuracy ML models (R2 >0.92) enabled real-time monitoring of glucose, lactate, viable cell density, and antibody titre. The glucose model was integrated into an automated process analytical technology (PAT) control strategy to maintain glucose at a predefined setpoint. The PAT strategy was compared to a manual bolus glucose control under high and low glucose feeding regimes. The PAT control approach observed an increase in biotherapeutic product titre by up to 35% and reduced glycation by up to 27%. These results indicated that both glucose setpoints and fluctuations impacted cell culture performance. In conclusion, this study highlighted the value of PAT tools in automating control loops, consequently improving process performance with enhanced productivity and quality.
In biopharmaceutical manufacturing, protein aggregation is a critical quality attribute, necessitating rapid and reliable analytical strategies during downstream processes like anion-exchange chromatography (AEX). While Raman spectroscopy enables continuous monitoring of protein secondary structure, standard data-driven regression models struggle to decouple intrinsic structural changes from gradient-induced solvent and buffer drifts under dynamic chromatographic conditions. Addressing this methodological gap, this study establishes a constrained pseudo-Voigt hard modeling framework for the mechanistic deconvolution of bovine serum albumin (BSA) size variants during in-line Raman monitoring of AEX processes. By explicitly defining a parametric background model to capture salt-induced spectral drift, the methodology effectively isolates matrix variations from genuine protein-specific signals. The constrained hard model was applied to 285 in-line spectra across diverse chromatographic conditions, achieving reconstruction fidelity while maintaining stable, physically interpretable component identities. The mechanistically derived Amide I center of mass emerged as a robust, aggregation-sensitive descriptor that preserves structural information despite strong concentration dynamics. Furthermore, the extracted spectral features demonstrated strong predictive performance for monomer concentration and acceptable accuracy for high molecular weight components. Collectively, these results demonstrate that constrained spectral hard modeling provides a highly interpretable, robust, and calibration-light alternative to classical partial least squares approaches for the real-time monitoring of protein size variants.
Sixty years of adeno-associated virus (AAV) research illustrates a trajectory marked by basic science exploration, iterative innovation, persistent challenges, a number of clinical setbacks, as well as commercial therapeutic triumphs. This continual evolution has led to recombinant AAV (rAAV) becoming a cornerstone of modern gene therapy. Significant advancements in molecular design, process development, and manufacturing have been made over the past three decades; these improvements are expected to significantly improve the safety, efficacy, and economics of rAAV gene therapies. Beyond rare disease, rAAV vectors have the potential to be used in prevalent conditions such as arthritis, heart failure, diabetes, Alzheimer's disease, and Parkinson's disease. Meeting the vector demands of these disease treatments will require continued innovations in rAAV manufacturing, including further improvements in process optimization and molecular engineering. In addition, the adoption of process intensification and automation strategies, pioneered in other biologics such as monoclonal antibody manufacturing, should prove pivotal in advancing the scale, robustness, and efficiency of rAAV production.
Rising global meat demand and nutritional awareness have fuelled interest in sustainable, ethical protein sources. Animal agriculture generates greenhouse gas emissions, land degradation, and water scarcity, creating a need for plant-based meat alternatives. While plant sources face drawbacks such as incomplete amino acid profiles, anti-nutritional factors, and land requirements. Algae emerge as a superior option, delivering exceptionally high protein content (up to 70% dry weight), complete essential amino acids, omega-3 fatty acids, vitamins, polysaccharides, and potent antioxidants, surpassing plant sources in nutrient density, bioavailability, and environmental footprint. This review evaluates the nutritional, environmental, and technological potential of key algal species (microalgae and macroalgae) for meat substitute applications. Algal formulations excel over plant-based counterparts with superior protein quality (PDCAAS >0.9 vs. often <0.8 for plants), rapid biomass growth (10-50× faster than plants), and no arable land requirements, enabling scalable, low-water production. The review addresses challenges such as off-flavors, digestibility, and cost through solutions such as strain selection, biorefinery optimization, and hybrid cultivation systems. An overview of key market players highlights the growing role of algae in alternative meats. By integrating nutritional and industrial perspectives, this work reveals trends positioning algae at the forefront for health-conscious consumers, advocating a "best-of-everything" approach with diverse species to revolutionize sustainable food systems.
Traditional off-line analysis methods to quantify residual metabolite concentrations in a drug substance fermentation process consume valuable time, require costly resources, and demand potentially hazardous manual operations when working with pathogenic biological organisms. This case study focuses on the successful method development and validation of Raman spectroscopy as an in-line process analytical technology (PAT) to replace the existing off-line assay for real-time glucose quantification. The value and feasibility of Raman spectroscopy as an in situ PAT method are recognized and demonstrated as an enabling technology suited to mitigate challenges fundamental to lab-scale process development and commercial manufacturing. Herein, application of chemometric techniques and multivariate analysis of Raman spectral data is described to develop a partial least squares model capable of predicting real-time and accurate glucose concentrations with a root mean square error of prediction of 1.2 g/L. Through this work, the predictive Raman PAT glucose quantification method was validated to make process decisions in a commercially licensed Pneumococcal vaccine drug substance manufacturing facility for the first time at our company. The method was integral to the process control strategy and applied to accurately and robustly indicate the glucose concentration target ± 2 g/L for triggering subsequent process operations. Several advantages were realized through this work including advanced process control, reduced operating costs, and improved safety posture.
Messenger RNA (mRNA) therapeutics can be used to protect against infectious disease and for the treatment of cancers and genetic disorders. In order to obtain the desired therapeutic dosage, it is necessary to concentrate the mRNA to levels where the viscosity of the mRNA solutions can become significant. However, there is currently no quantitative data on the viscosity of these concentrated mRNA solutions. Experiments were performed with two model mRNA constructs: a firefly luciferase (Fluc) mRNA (2.117 kb) and a self-amplifying RNA (9.442 kb) over a wide range of concentrations. The viscosity of the larger saRNA was more than 140 mPa·s at a concentration of 28 mg/mL, which is more than 10× larger than that for the Fluc mRNA under the same conditions. The viscosity was a weak function of shear rate (from 10 to 100 s-1), with some shear-thinning behavior. The viscosity data were analyzed using several available models from the literature, providing additional insights into the underlying physical behavior. The potential implications of the mRNA viscosity in the downstream processing of mRNA therapeutics are also discussed.
Endemic human coronaviruses OC43 and HKU1 cause widespread respiratory infections and can be associated with severe illness in immunocompromised and elderly individuals. Frequent adaptive evolution in the spike proteins of these embecoviruses and the potential for zoonotic transmission from a large animal reservoir necessitates the characterization of the immunogenic landscape of the spike proteins of embecoviruses. Here, we constructed nanoparticle vaccines displaying the spike antigens from OC43, HKU1 A, or HKU1 B, as well as a bivalent formulation incorporating spike antigens from OC43 and HKU1 A. Immunization of mice elicited spike-specific IgG antibody responses, with endpoint titers demonstrating cross-reactivity among clade-matched viruses. Notably, the bivalent formulation elicited antibody responses comparable to those of monovalent vaccines against matched antigens. These findings inform future design of vaccines against human-infecting embecoviruses and could serve as an important step toward a universal vaccine against common cold causing coronaviruses.
The rapidly developing antibody market poses challenges to the production cost and efficiency, which can be addressed through continuous process. However, a hurdle of high capital investment remains a concern especially for the company with existing fed-batch capacity. This study proposes a straightforward approach to convert existing batch production suites into continuous ones using membrane-based technology. A conceptual validation was conducted at a 100 L pilot scale, and a head-to-head comparison was carried out. The results demonstrated that Protein A membrane chromatography could achieve a productivity 6.3 times higher than that of periodic counter-current chromatography (PCC), without requiring the purchase of any dedicated equipment. Using Q membrane, compared with traditional column anion exchange chromatography, can achieve a 16-fold increase in load capacity and a 25-fold increase in productivity. Finally, VF-UF/DF (Virus Filtration-Ultrafiltration/diafiltration) linkage process was designed, resulting in a 20% reduction in process time. During the 23-day upstream perfusion and 15-day downstream processing, the cumulative titer reached 51.0 g/L, with an 83.1% yield in downstream processing. With downstream process time shortening from 4 to 1.5 days, the process capacity could be improved by 8 times using the existing batch production suites and the same facility area. PATs, including ultra-performance liquid chromatography, Raman spectroscopy, FlowVPX, and an inline turbidity meter, were integrated to monitor process CQAs (Critical Quality Attributes): monomer purity, protein concentration, and turbidity, to enable efficient process control in the future.
Endotoxins pose a significant risk to drug product safety due to their potent pyrogenic activity. However, developing a universal decontamination method for protein solutions remains challenging, as current technologies struggle to reconcile efficient endotoxin removal with high product recovery and operational simplicity. This study explores the novel application of commercial virus filters for endotoxin clearance. We systematically evaluated filters from multiple manufacturers across a range of initial endotoxin concentrations and buffer conditions. The results demonstrate that Planova BioEX and Valpha PES virus filters reduced endotoxin levels by 88%-99% at initial endotoxin concentrations below 5 EU/mg of a therapeutic monoclonal antibody, while achieving near-complete protein recovery (>95%). A comparative study revealed that under low pH conditions (pH 5.5), virus filters outperformed Mustang E and Mustang Q membranes in removing low levels of endotoxin. Under high pH conditions (pH 7.5), their performance was comparable. This difference likely stems from distinct removal mechanisms: Mustang membranes rely on electrostatic interactions, whereas virus filters (~20 nm pore size) operate via size exclusion. Dynamic light scattering results indicated the majority of the mAb-endotoxin complexes exceeded 20 nm, supporting the size-based removal mechanism of virus filters. These findings demonstrate that virus filtration has the potential to serve as a viable alternative for endotoxin removal in preclinical protein processing, particularly for high-pH sensitive proteins and acidic proteins. It also represents a valuable addition to the endotoxin clearance toolbox for clinical manufacturing.
