Biotechnol. Prog. [JOURNAL]
Sun
200 papers
Enhanced lentiviral vector recovery and separation using arginine hydrochloride with CIM QA monoliths.
Kocot AJ, Kulkarni S, Ghosh R … +4 more , Altern SH, Dordick JS, Przybycien TM, Cramer SM
Biotechnol Prog · 2026 · PMID 41572492 · Publisher ↗
Lentiviral vectors (LVVs) offer distinct advantages including large payload capacity and stable transduction of non-dividing cells, making them well-suited for ex vivo modification of stem or immune cells. However, chrom...
Lentiviral vectors (LVVs) offer distinct advantages including large payload capacity and stable transduction of non-dividing cells, making them well-suited for ex vivo modification of stem or immune cells. However, chromatographic purification of LVVs is hindered by low recoveries, co-elution of product related impurities, and vector instability. In this study, we evaluated arginine hydrochloride (ArgHCl) as an alternative eluent to sodium chloride using CIMmultus™ QA (CIM QA) monoliths. Screening of solution conditions identified that phosphate buffer concentrations between 100-200 mM enhanced infective particle stability. During anion exchange screening experiments using a CIM QA 96-well plate, ArgHCl improved both infectious particle and p24 recoveries, which was corroborated in linear gradient elution (LGE) experiments using the monolith in a disk format. Furthermore, fractions eluted with ArgHCl exhibited improved colloidal stability compared to those eluted with NaCl. Increasing the ArgHCl gradient endpoint concentration to 1.5 M ArgHCl yielded infectious particle recoveries of 71%. Analysis of gradient fractions with nanoflow cytometry revealed a two-peak elution profile, with the more strongly retained peak enriched in vesicular stomatitis virus glycoprotein (VSV-G) positive particles, corresponding to greater infectious particle recoveries. ArgHCl also improved the chromatographic resolution between the initial impurity peak and the secondary peak, enabling improved separation. These findings support the use of ArgHCl and phosphate buffers to enhance recovery during the purification of LVVs using AEX chromatography and highlight the utility of nanoflow cytometry for rapid detection of product-related impurities.
Unveiling microbial and microalgal chassis for therapeutic and diagnostic protein expression.
Bhandari L, Kulkarni S, Prakash G
Biotechnol Prog · 2026 · PMID 41562205 · Publisher ↗
Biopharmaceuticals are becoming one of the most successful clinical therapeutic products for treating various disorders and are gradually being utilized across nearly all areas of medicine. They have revolutionized the t...
Biopharmaceuticals are becoming one of the most successful clinical therapeutic products for treating various disorders and are gradually being utilized across nearly all areas of medicine. They have revolutionized the treatment of numerous diseases and continue to represent a significant area of research and development. Presently, host cell systems like bacteria, yeast, insects, and mammalian cells dominate the production of both therapeutic and diagnostic proteins. This review explores the strengths and limitations of these existing host systems for recombinant protein production, emphasizing the promising potential of microalgal systems for expressing therapeutic and diagnostic proteins. It accentuates the advantages of microalgae, such as their rapid growth rates, scalability, and sustainability. We delve into the intricacies of glycosylation patterns in microalgae, comparing them with those in other expression systems. This review highlights recent advancements in algal-based protein expression systems for diagnostic and therapeutic applications. It also outlines a strategic roadmap for future developments in biopharmaceutical production, emphasizing how each expression system's unique characteristics can help meet modern medicine's growing demands.
A size-exclusion chromatography fingerprinting workflow for the development of flow-through polishing operations for mAbs derived from continuous precipitation processes.
Gutierrez-Diaz MA, Altern SH, Przybycien TM … +1 more , Cramer SM
Biotechnol Prog · 2026 · PMID 41552891 · Publisher ↗
In this work we present a workflow for developing two-step, flow-through polishing processes for monoclonal antibodies (mAbs). The approach is demonstrated using redissolved precipitates from three CHO-derived mAbs gener...
In this work we present a workflow for developing two-step, flow-through polishing processes for monoclonal antibodies (mAbs). The approach is demonstrated using redissolved precipitates from three CHO-derived mAbs generated by a continuous, PEG/ZnCl₂-mediated precipitation capture process. Size Exclusion Chromatography (SEC) fingerprinting and a percent SEC clearance (PSC) metric are developed to enable simultaneous quantification of monomer yield and impurity removal during high-throughput screening and scale-down column studies. Batch slurry plate screens are used to evaluate multimodal anion exchange (MMA) resins and an activated carbon composite adsorber under varying pH and ionic strengths, assessing partition coefficients and PSC values against both low-molecular-weight (LMW) and high-molecular-weight (HMW) impurities. Top candidates were then assessed in single-column, higher-loading flow-through experiments using the redissolved precipitates as feeds. Activated carbon emerged as a highly effective first polishing step for LMW impurity removal under acidic, low-conductivity conditions, while MMA resins provided complementary LMW and HMW clearances in a subsequent flow-through step. The two-step processes achieved overall mAb recoveries of 80%-87%, reduced HMW species from >1.7% down to 1.1%, and decreased host-cell protein levels from >10,000 to <40 ppm for all three mAbs. SEC fingerprints showed the ability to identify orthogonal impurity removal opportunities between the two polishing materials, validating the screening methodology for a process devoid of bind-elute processing steps. This work demonstrates that SEC-based impurity profiling and PSC metrics can guide the development of flow-through polishing processes and offer a useful intensification strategy to alleviate DSP bottlenecks and reduce reliance on affinity capture.
Development of a co-culture of Ureibacillus thermosphaericus and Cupriavidus taiwanensis for inhibitors removal from hemicellulose prehydrolysate.
Theiri M, Marinova M, Chadjaa H … +1 more , Jolicoeur M
Biotechnol Prog · 2026 · PMID 41552861 · Full text
For biofuels production, hemicellulose pre-hydrolysate is considered an attractive feedstock rich in fermentable sugars. The lignocellulosic biomass comprises, along with sugars, several inhibitors that can hamper its ef...
For biofuels production, hemicellulose pre-hydrolysate is considered an attractive feedstock rich in fermentable sugars. The lignocellulosic biomass comprises, along with sugars, several inhibitors that can hamper its efficient conversion. In this work, mixed cultures of Ureibacillus thermosphaericus and Cupriavidus taiwanensis were used for the first time to detoxify the pre-hydrolysate. The nutrient source was first optimized in synthetic media with mono-cultures to detoxify phenolic compounds, and a medium containing inorganic salts was selected. Afterwards, the efficiency of phenolic degradation was compared in a single-compound solution and in a mixture. The simultaneous co-culture showed the highest degradation efficiency (90% at 2.8 g/L of phenolic compounds). Finally, the detoxification of a raw pre-hydrolysate was conducted, and a maximum degradation of 14% of the phenolics was obtained using sequential inoculation of Ureibacillus thermosphaericus followed by Cupriavidus taiwanensis addition.
A case study showing the role of hydrophobicity variants and other enriched mAb proteoforms on filterability through a virus filter with productivity improvement measures.
Isu S, Silva D, Holstein M … +4 more , Lewandowski A, Cunningham K, Sokolnicki A, Raghunath B
Biotechnol Prog · 2026 · PMID 41549894 · Full text
A rapid assessment of manufacturability for drug candidates is crucial for advancing a prospective biotherapeutic from a candidate to a bulk drug substance. A lot-to-lot approach to manufacturability is adopted where eac...
A rapid assessment of manufacturability for drug candidates is crucial for advancing a prospective biotherapeutic from a candidate to a bulk drug substance. A lot-to-lot approach to manufacturability is adopted where each biologic batch is assessed for manufacturability as a bulk, unfractionated pool. Manufacturers may explore a more granular approach, independently enriching and evaluating the filterability of antibody variants within each lot, especially within the confines of relative hydrophobicity and surface charge. This study examined the use of bind-and-elute chromatography to alter the proportions of monoclonal antibody (mAb) proteoforms in eluate sub-pools from a mixed-mode chromatography resin-packed column. Filterability of each sub-pool through a virus-retaining filter was subsequently examined. Circular dichroism and Fourier transform infrared spectroscopy were performed for each sub-pool to probe for higher-order structure differences between mAb variants enriched therein. Bioanalytical techniques were also used to assess colloidal stability, surface hydrophobicity, surface charge, and size differences. Results showed that basic charge variants, high-mannose glycovariants, high relative hydrophobicity proteoforms, and high-molecular-weight species were enriched in the last-eluting (terminal) sub-pools. The first sub-pool and the final sub-pool showed the most fouling propensity on VPro virus filters. Circular dichroism showed that enriched proteoforms in the last sub-pool possessed a higher percentage of bends. Most secondary structures did not vary significantly between sub-pools. Diffusion interaction parameter was highly negative across all sub-pools and the bulk unfractionated pool. These results provide a design space for identifying and depleting problematic mAb variants before the crucial virus filtration step.
Performance of large virus removal filters during AAV processing: Influence of flux and process disruptions.
Chaubal AS, Arachchige AW, Zahn AJ … +3 more , Wickramasinghe SR, Qian X, Zydney AL
Biotechnol Prog · 2026 · PMID 41543236 · Full text
As adeno-associated viral vectors (AAV) continue to advance through the clinical pipeline, effective downstream purification strategies must be developed to ensure bulk drug purity and safety. AAV are produced within mam...
As adeno-associated viral vectors (AAV) continue to advance through the clinical pipeline, effective downstream purification strategies must be developed to ensure bulk drug purity and safety. AAV are produced within mammalian cells, bringing forth risks associated with viral contamination. Although existing downstream operations provide some degree of viral inactivation and removal, regulatory agencies have recommended the incorporation of a dedicated virus removal filtration step to ensure robust viral clearance. Recently published studies have demonstrated that membrane filters with nominal pore sizes between 35 and 50 nm can provide effective AAV transmission while removing larger viruses, although these results were obtained over a limited range of conditions. This study represents the first investigation into the effects of filtrate flux and process disruptions on virus reduction filtration for AAV. Experiments were performed using purified AAV capsids and carboxylate-modified polymeric nanoparticles with a nominal diameter of 20 nm. Initial results confirmed that both systems exhibited nearly identical transient transmission profiles during virus filtration. Virus filtration performed at various filtrate fluxes (between 20 and 185 L/m/h) revealed that moderately higher AAV yield may be obtained at lower fluxes. The data were analyzed using a modified internal polarization model, which was extended to account for the effects of process disruptions on transient particle transmission and recovery. Process disruptions were employed to increase AAV yield beyond 99% without compromising overall clearance of large viruses. At least a 4-log reduction in xenotropic murine leukemia virus (XMuLV) was observed under all conditions tested, even following multiple process pauses.
Dynamics of natural killer cell function upon recurrent stimulation.
One J, Narayan J, Cichocki F … +2 more , Hu WS, Azarin SM
Biotechnol Prog · 2026 · PMID 41531166 · Full text
Natural killer (NK) cells have shown potential for allogeneic cell-based cancer immunotherapies. For development of economical off-the-shelf allogeneic therapies, maximal expansion of the NK cells from each donor must be...
Natural killer (NK) cells have shown potential for allogeneic cell-based cancer immunotherapies. For development of economical off-the-shelf allogeneic therapies, maximal expansion of the NK cells from each donor must be achieved while maintaining efficacy and uniformity of the cell product. The standard method for robust expansion utilizes weekly stimulation with engineered feeder cells derived from the K562 cell line. However, the effects of repeated stimulation on NK cell growth, metabolism, and function are not well understood. In this study, we demonstrated a distinct shift in growth kinetics and metabolism around week 3-4 of repeated K562 feeder cell stimulation, followed by a change in cytokine secretion and killing ability. Seahorse metabolic flux assays and transcriptomics suggested a transition from glycolytic metabolism to oxidative metabolism after the first week of stimulation, but the shift in growth kinetics generally correlated to reduced metabolic activity. Collectively, these results indicate that serial stimulation sustains large-fold NK cell expansion that can be exploited for NK cell therapy; however, this expansion has important impacts on NK cell growth, metabolism, and function. Careful characterization is critical when developing large-scale biomanufacturing processes to ensure efficacy of the final cellular product.
Design considerations impacting flow dynamics in packed beds for virus inactivation.
Basria I, Ajayi O, FuentesArias M … +2 more , Najarro AO, Lute S
Biotechnol Prog · 2026 · PMID 41518105 · Publisher ↗
Despite significant advances in continuous manufacturing of monoclonal antibodies, the implementation of continuous virus inactivation (CVI) remains challenging due to standardization gaps that could compromise product q...
Despite significant advances in continuous manufacturing of monoclonal antibodies, the implementation of continuous virus inactivation (CVI) remains challenging due to standardization gaps that could compromise product quality and safety. This study identified limitations in minimum residence time (mRT) prediction for packed bed reactors (PBR) utilized for CVI. This work focused on characterizing the residence time distribution (RTD) behavior of tracers with varying molecular properties in four PBR configurations. The results demonstrated that tracer molecular size impacted mRT prediction, with larger molecules showing shorter residence times than smaller molecule tracers under identical conditions. During scale-up from 16 to 26 mm diameter columns, mRT was not maintained, suggesting that traditional chromatography scale-up principles may not be directly applicable to CVI using PBRs. Overall, this work established a helpful foundational understanding of how process material properties impact mRT prediction-a critical process parameter that would directly impact virus inactivation efficacy in integrated CVI systems.
Utilizing Cole-Cole parameters for in-line feedback: Cell culture process adjustments based on cell health.
Krishnan N, Jiang T, Crowe J … +6 more , Gotta S, Wittmer T, Staffenhagen J, Alvarado B, Jambunathan P, Rameez S
Biotechnol Prog · 2026 · PMID 41457296 · Publisher ↗
In the biopharmaceutical industry, effective process control strategies are essential for enhancing drug substance quality and yield. This study presents dielectric spectroscopy as a novel approach for in-line monitoring...
In the biopharmaceutical industry, effective process control strategies are essential for enhancing drug substance quality and yield. This study presents dielectric spectroscopy as a novel approach for in-line monitoring of cell apoptosis, enabling earlier detection of apoptotic events and related cellular changes. Utilizing permittivity measurements, we assessed Cole-Cole parameters, specifically critical frequency (f) and delta epsilon (Δε), as key performance indicators (KPIs) for real-time monitoring of cell health in CHO cell cultures during batch and perfusion processes. Our findings demonstrate that variations in these parameters correlate with cellular stress responses, such as nutrient limitation and high shear conditions, providing timely signals for process monitoring and control. By integrating these in-line measurements, we can enhance feeding strategies, ultimately improving cell viability and productivity. This approach not only streamlines the monitoring process but also offers a robust framework for proactive adjustments in bioprocessing, thereby optimizing overall performance and resource utilization.
Purification of antisense oligonucleotides using hydrophobic interaction chromatography.
Gronke RS, Immel-Brown JP, Jeyabalan S … +5 more , Banzon PD, Delavari A, Tello JC, Joshi R, Ho T
Biotechnol Prog · 2026 · PMID 41449859 · Publisher ↗
Hydrophobic interaction chromatography (HIC) provides a powerful alternative impurity control method for antisense oligonucleotide purification relative to traditionally used anion exchange (AEX) and/or reverse phase met...
Hydrophobic interaction chromatography (HIC) provides a powerful alternative impurity control method for antisense oligonucleotide purification relative to traditionally used anion exchange (AEX) and/or reverse phase methods. HIC is particularly effective in clearing process-related solvents and small molecules by ≥3 log as well as failure sequences (sometimes called early eluting impurities (EEIs) by ≥90%). Additionally, HIC reduces harder to remove product-related impurities. These include branchmers (late eluting impurities (LEIs), oligonucleotides missing a single nucleotide (N-1 impurities), oligonucleotides lacking appropriate phosphorothioate sulfurization (P = O impurity), and other synthesis-related impurities. To optimize the purification process, variables such as resin ligand, salt types, processing conditions, types of gradients, and loading ratios were systematically evaluated to achieve 90% yield and maximal impurity resolution. Loading the column at 32%-78% of its dynamic binding capacity (DBC), combined with stepwise wash and elution gradients, provided effective resolution of impurities in crude oligonucleotide mixtures. The desorption of the purified product was achieved in low lyotropic salt concentrations (typically ≤50 mM) using a stepwise gradient. This approach retained non-polar impurities such as LEIs within the column. When properly designed, HIC is an all-aqueous, scalable, cost effective and predictable purification process. It can be implemented as a stand-alone method or integrated into a dual-column process alongside orthogonal techniques, such as AEX, to achieve even higher levels of product purity.
In silico mediated development of orthogonally selective mAb downstream processes for the removal of process-related impurities.
Jang D, Gutierrez-Diaz MA, Altern SH … +2 more , Tjandra H, Cramer SM
Biotechnol Prog · 2026 · PMID 41400331 · Publisher ↗
Continued advancements in recombinant CHO expression of therapeutic mAbs have led to improved productivity but have also increased the HCP burden on the downstream purification process. In this work, we developed an in s...
Continued advancements in recombinant CHO expression of therapeutic mAbs have led to improved productivity but have also increased the HCP burden on the downstream purification process. In this work, we developed an in silico mediated workflow to facilitate the rapid development of non-protein A three-step processes for the effective removal of HCPs from a CHO-derived mAb therapeutic. Null CCF and pure mAb retention patterns were generated using linear gradient screens on a set of strategically selected resins, membrane adsorbers, and novel adsorbents. HCP characterization of key fractions was then carried out using RPLC "HCP fingerprinting" and the resulting retention database was processed using an in silico tool to generate a list of all possible three-step sequences subject to design constraints. Top-ranked processes generated by the tool were then evaluated and refined at the bench scale to produce several successful processes consisting of bind-elute capture followed by either a bind-elute and flowthrough step (91.4 ppm HCP with a cumulative product yield of 78.7%) or two flowthrough steps with no salt (96.1 ppm HCP with a cumulative yield of 81.4%).
Enhancing CHO cell recombinant protein production using a perfusion-directed host evolution approach.
Amaya P, Mistry RK, Sou S … +5 more , Hess S, Khanal B, Schaeffer Z, Zhu J, Chakrabarti L
Biotechnol Prog · 2026 · PMID 41307185 · Full text
Clonally derived cell lines generated from Chinese hamster ovary (CHO) cells encounter numerous stressors when cultured in high-intensity perfusion bioreactors leading to poor process performance. To circumvent this, the...
Clonally derived cell lines generated from Chinese hamster ovary (CHO) cells encounter numerous stressors when cultured in high-intensity perfusion bioreactors leading to poor process performance. To circumvent this, the ability of CHO cells to adapt to different culture environments was exploited. Here host cells were selected in the presence of physical and chemical stressors associated with a perfusion environment by culturing at a high cell density in a perfusion bioreactor for 30 days. Following recovery and expansion, the performance of the resulting perfusion-evolved host was evaluated using stable transfectant pools and clones expressing biotherapeutics of different formats. Cell lines generated from the perfusion host outperformed the parental host at several fundamental stages of the clone selection process. Perfusion host-derived pools showed elevations in productivity, cell-specific productivity, end-of-run viability, and reduced lactate production in fed-batch culture. Use of the perfusion host for cell line generation resulted in an increased frequency of high-producing clones. Moreover, the perfusion host-derived clones demonstrated 30% higher productivity and improved mannose profile in the perfusion environment compared to the clones from the parental host. Furthermore, a comparative proteomic analysis between the two host types revealed unique regulatory networks that allowed us to gain insights into the underlying molecular processes influencing production performance. Taken together, the results suggest that the perfusion host may not only increase the efficiency of the cell line development process but may also serve as an efficient tool for improvement in production capability in the perfusion platform.
Analysis of the economic viability and environmental impacts of a conceptual process model for the recovery of lactic acid from spent media in cultivated meat production.
Wimble J, Ashizawa R, Swartz EW
Biotechnol Prog · 2026 · PMID 41273040 · Full text
Scaled production of cultivated meat (CM) will co-produce large volumes of spent media. Recycling of abundant metabolites such as lactic acid in spent media offers an opportunity for valorization and reduction of the car...
Scaled production of cultivated meat (CM) will co-produce large volumes of spent media. Recycling of abundant metabolites such as lactic acid in spent media offers an opportunity for valorization and reduction of the carbon footprint of CM production; however, the feasibility has yet to be examined. We modeled a conceptual five-step lactic acid recovery process integrated into a previously modeled CM facility and analyzed the corresponding cost and environmental impacts of recovering an 88% lactic acid solution. At an anticipated lactic acid concentration in spent media of 3 g/L, we found the net cost of recovery would be $0.71/kg lactic acid, with a 7.5-year simple payback period. Sales of lactic acid as a co-product could offset $0.06/kg of the cost of CM production. Depending on allocation scenarios, the environmental impact of CM production with an integrated recovery process had a -1.0 to +0.2 kg CO eq effect on the carbon footprint and a -22 to +3 MJ effect on cumulative energy demand per kg of CM. Recovery of lactic acid from spent media also had a 25% lower carbon footprint than conventional fermentation processes. These model results suggest that recovery of lactic acid may be an economically viable and environmentally beneficial practice if validated in future CM production facilities. This original study provides crucial guidance for lactic acid valorization and other media recycling strategies that can be broadly applied to animal cell biomanufacturing industries.
Effect of cell retention techniques in Komagataella phaffii lab-scale continuous processes.
Linova MY, Kodiripaka SK, Martins E … +3 more , Sripada SA, Menegatti S, Woodley JM
Biotechnol Prog · 2026 · PMID 41255079 · Full text
Perfusion technologies play a growing role in the implementation of continuous processes for biotherapeutics production in mammalian-based manufacturing. However, their application to alternative production hosts is limi...
Perfusion technologies play a growing role in the implementation of continuous processes for biotherapeutics production in mammalian-based manufacturing. However, their application to alternative production hosts is limited. Cell retention systems are of key importance for the efficiency of perfusion bioreactors. In this study, we investigate two cell retention technologies for the development of lab-scale Komagataella phaffii continuous processes. An acoustic-based process (AP) and a membrane-based process (MP) were developed using an acoustic cell separator (ACS) and a vibrating membrane filtration (VMF) device, respectively. Both systems allowed for continuous cell recycle and production of scFv13R4 antibody fragment for 8 days (AP) and 9 days (MP), without loss in productivity, while maintaining high viability (greater than 90%). Higher volumetric and specific productivities were achieved during the AP process, namely 50.63 ± 1.63 mg L day and 1.09 ± 0.07 mg g day, against the 32.29 ± 1.21 mg L day and 0.44 ± 0.02 mg g day afforded by the MP process. The VMF device provided 100% separation efficiency with biomass accumulating up to concentrations of 74.1 ± 0.1 g L dry cell weight (DCW), whereas the acoustic device reached 55.1 ± 0.47 g L DCW at 98% separation efficiency. The acoustic device showed selectivity towards larger and more complex cells in the yeast population, which might be linked to the observed higher productivities for the AP process. This study discusses the advantages and drawbacks of both cell retention technologies and provides an outlook towards their future investigation in K. phaffii perfusion processes.
Augmenting therapeutic protein production in CHO cells: A proline-based selection strategy for enhanced productivity and product quality.
Zhao B, Zhang B, Kang Y … +8 more , Liu S, Jia Z, Lu C, Cao Y, Xu A, Lee KS, Zhang Z, Song J
Biotechnol Prog · 2026 · PMID 41249774 · Publisher ↗
Chinese hamster ovary (CHO) cells have emerged as the predominant mammalian host for the production of therapeutic recombinant proteins, including monoclonal antibodies (mAbs), bispecific antibodies (bsAbs), and fusion p...
Chinese hamster ovary (CHO) cells have emerged as the predominant mammalian host for the production of therapeutic recombinant proteins, including monoclonal antibodies (mAbs), bispecific antibodies (bsAbs), and fusion proteins. To meet the growing demand for biologics and reduce manufacturing costs, the exploitation of efficient cell line development platforms is essential. Over the past decades, various selection markers, such as dihydrofolate reductase (DHFR), glutamine synthetase (GS), and antibiotic resistance genes, have been widely utilized in the development of production cell lines. In this study, we introduce the proline selection system, an alternative metabolic selection strategy, as an efficient approach to optimize our CHO cell line development platform. By employing yeast PRO1 and PRO2 genes as selection markers, proline selection effectively complements GS selection to establish high-producing cell lines for both mAbs and bsAbs. In particular, the integration of PRO1 and PRO2 genes into a single plasmid, in conjunction with the GS gene, significantly enhances productivity for asymmetric molecules. Optimized chain configuration across proline and GS selection plasmids can further boost protein yield. Additionally, the overexpression of regulator proteins can be leveraged with proline selection to enhance antibody production or fine-tune product quality. Taken together, the incorporation of proline selection into CHO cell line development, particularly when combined with GS selection, provides a consistent and streamlined strategy to meet the growing demand for high-quality biologics in the pharmaceutical industry.
Image analysis method for measurement and prediction of intra-matrix IgG diffusion.
Debbarma R, Dos Santos ACF, Ladisch M
Biotechnol Prog · 2026 · PMID 41243619 · Full text
Measurement and imaging of intra-matrix protein therapeutics diffusion is important due to the emergence of injectable biologics currently in various stages of research and clinical testing. These therapeutics are develo...
Measurement and imaging of intra-matrix protein therapeutics diffusion is important due to the emergence of injectable biologics currently in various stages of research and clinical testing. These therapeutics are developed for delivery to hyaluronic acid (HA)-rich anatomical sites such as subcutaneous tissue, the vitreous humor, and knee joints, depending on the target tissue. Understanding their diffusion behavior is essential for optimizing drug delivery strategies. Our work presents an image analysis method suited for tracking IgG diffusion in low viscosity HA matrices representative of the vitreous humor, where diffusion occurs more rapidly unlike a previously reported analysis method for higher viscosity matrices where protein diffusion is significantly slower. The current method utilizes scanner images at 6.3 MP resolution, and an algorithm that removes background and calculates protein mass and concentration measured directly within matrices formulated to represent HA in an intravitreal environment. We report and demonstrate a robust method for predicting protein diffusion coefficient from images of label-free protein diffusing in a low viscosity HA matrix.
RETRACTION: Comparison of Expression Optimization of New Derivative of staphylokinase (SAK-2RGD-TTI) with the rSAK.
Biotechnol Prog · 2025 · PMID 41215551 · Publisher ↗
H. Faraji , M. Ramezani , B. Mashkani , H. R. Sadeghnia , H. M. Benhangi , S. H. Teshnizi , and F. Soltani , "Comparison of Expression Optimization of New Derivative of staphylokinase (SAK-2RGD-TTI) with the rSAK," Biote...
H. Faraji , M. Ramezani , B. Mashkani , H. R. Sadeghnia , H. M. Benhangi , S. H. Teshnizi , and F. Soltani , "Comparison of Expression Optimization of New Derivative of staphylokinase (SAK-2RGD-TTI) with the rSAK," Biotechnology Progress 35, no. 4 (2019): e2819. 10.1002/btpr.2819. The above article, published online on 11 April 2019 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between journal Editor-in-Chief, John A. Morgan; American Institute of Chemical Engineers, and Wiley Periodicals, LLC. A third party reported that Figure 7 contained several repeated image elements and that a number of these elements were copied from a previous publication by some of the same authors (Faraji et al. 2017 [https://doi.org/10.1080/10826068.2016.1252924]). An investigation by the publisher confirmed these concerns and also found that the protein marker in Figure 9 had been copied from another publication (Pednekar et al 2016 [https://doi.org/10.3389/fimmu.2016.00567]) and that elements in Figure 11B had been duplicated and manipulated. The authors did not respond to an inquiry and request for original data by the publisher. The retraction has been agreed to because the evidence of image manipulation fundamentally compromises the editors' confidence in the results presented.
Artificial intelligence and machine learning-assisted digital applications for biopharmaceutical manufacturing.
Panjwani S, Wei H, Mason J
Biotechnol Prog · 2026 · PMID 41211798 · Full text
Artificial intelligence and automation are no longer just buzzwords in the biopharmaceutical industry. The manufacturing of a class of biologics, comprising monoclonal antibodies, cell therapies, and gene therapies, is f...
Artificial intelligence and automation are no longer just buzzwords in the biopharmaceutical industry. The manufacturing of a class of biologics, comprising monoclonal antibodies, cell therapies, and gene therapies, is far more complex than that of traditional small molecule drugs. Therefore, applications based on artificial intelligence are essential for successfully manufacturing this new class of biologics more quickly and more economically. Some biologics manufacturers, academic researchers, and young entrepreneurs have already begun implementing artificial intelligence-based applications to increase operational efficiency, enhance process understanding, improve process monitoring, and achieve better regulatory compliance. Regulatory guidance from health agencies on the use of artificial intelligence and machine learning is acting as a catalyst in the adoption process of these new technologies by the biopharmaceutical industry. Research in artificial intelligence and machine learning has also advanced significantly in the last decade. At the same time, new cloud technologies have made the development and deployment of machine learning applications much easier. Several examples of artificial intelligence and machine learning applications in monoclonal antibodies manufacturing already exist. Cell and gene therapy, which present the future of medicine, will also benefit from this new technology. Overall, advancements in this domain will essentially help better serve patients' needs.
Optimizing sterile filtration of nanoemulsions through proper choice of prefilter properties.
Kapila S, Soukup RJ, Bradley ME … +2 more , Boyd D, Zydney AL
Biotechnol Prog · 2026 · PMID 41170564 · Full text
Nanoemulsions, with droplet sizes between 20 and 200 nm, have emerged as a promising vaccine adjuvant and drug delivery system, enhancing the solubility of hydrophobic drugs for diverse applications. Sterile filtration o...
Nanoemulsions, with droplet sizes between 20 and 200 nm, have emerged as a promising vaccine adjuvant and drug delivery system, enhancing the solubility of hydrophobic drugs for diverse applications. Sterile filtration of nanoemulsions is particularly challenging due to the similar size between the nanodroplets and the 0.2 μm nominal pore size rating of sterile filters. One approach to reducing membrane fouling, and enhancing filtration capacity and yield, is to employ an appropriate prefilter, but there are currently no clear guidelines on how to select the prefilter pore size, chemistry, or morphology for sterile filtration of nanoemulsions. This study examined the performance of a range of prefilters with varying pore morphologies and surface chemistries. Sessile drop contact angles were used to evaluate the prefilter hydrophobicity, and bubble point and mercury intrusion porosimetry were used to evaluate the pore characteristics of the different prefilters. The best performance was achieved using a relatively hydrophobic 0.45 μm prefilter made of polyvinylidene fluoride but modified with a somewhat hydrophilic (oxygen-containing) coating. This prefilter reduced the surface tension of the nanoemulsion and provided more than a 2-fold increase in capacity for a variety of sterile filters. These results provide critical insights into the factors influencing nanoemulsion filtration and offer a framework for selection of appropriate prefilters in biopharmaceutical manufacturing.
Kinetic and equilibrium analysis of electrochemical Aptasensing for real-time detection of Staphylococcus aureus in food substances.
Soleimani S, Bruce-Tagoe TA, Danquah MK
Biotechnol Prog · 2026 · PMID 41146495 · Publisher ↗
Real-time detection of foodborne pathogens such as Staphylococcus aureus (S. aureus) is essential for ensuring food safety. In this study, we evaluate the performance of an electrochemical aptasensor developed from gold...
Real-time detection of foodborne pathogens such as Staphylococcus aureus (S. aureus) is essential for ensuring food safety. In this study, we evaluate the performance of an electrochemical aptasensor developed from gold nanoparticles (AuNPs)-immobilized screen-printed carbon electrode for the detection of low concentrations of S. aureus in chicken extract media. Using cyclic voltammetry (CV), the dynamic interaction between the aptamer-modified electrode and S. aureus was monitored across four bacterial concentrations of 1, 5, 10, and 20 colony-forming units per milliliter (CFU/mL) at 35-min intervals over 350 min. The aptasensor demonstrated a concentration-dependent response with increasingly lower maximum CV signals and faster time to equilibrium as CFU increased. Real-time kinetic and equilibrium parameters were extracted to understand the binding behavior of the pathogen to the electrode surface. Critical parameters such as the kinetic rate constant (k) of 0.0274 min and equilibrium dissociation constants ( ) of 7.35 CFU/mL, were derived from the CV signals. Langmuir isotherm modeling yielded a maximum binding capacity ( ) of 33.55 μA. In addition, a Hill coefficient (n) of 0.65 was obtained, which indicates a slightly negative cooperativity. These findings demonstrate the capability of the aptasensor for real-time detection of S. aureus, offering a robust framework for field-deployable pathogen monitoring in food matrices.
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