Biotechnology Progress最新文献_第3页

Small scale purification and biophysical characterization of haptoglobin derived from human Cohn fraction IV. 人Cohn蛋白IV的小尺度纯化及生物物理特性研究。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-09-06 DOI: 10.1002/btpr.88555

Shuwei Lu, Mohd Asim Khan, Pedro Cabrales, David C Irwin, Paul W Buehler, Andre F Palmer

{"title":"Small scale purification and biophysical characterization of haptoglobin derived from human Cohn fraction IV.","authors":"Shuwei Lu, Mohd Asim Khan, Pedro Cabrales, David C Irwin, Paul W Buehler, Andre F Palmer","doi":"10.1002/btpr.88555","DOIUrl":"https://doi.org/10.1002/btpr.88555","url":null,"abstract":"Haptoglobin (Hp) is a polymorphic acute phase α-2 glycoprotein found in plasma that plays a critical role in binding, neutralizing, and removing cell-free hemoglobin (Hb) from the circulation. Under clinical conditions characterized by high levels of hemolysis, such as in patients with sickle cell disease (SCD), large quantities of cell-free Hb are released from lysed red blood cells (RBCs) into the circulation and bind to Hp. This interaction reduces the plasma Hp concentration below basal levels and diminishes its Hb-binding capacity. Therefore, plasma-derived Hp has the potential to be used therapeutically to scavenge, neutralize, and remove excess cell-free Hb from the blood, thus preventing Hb-mediated toxicity. This provides strong motivation to purify Hp at high purity using sustainable sources, such as waste plasma fractions from the Cohn plasma fractionation process used to produce human serum albumin from pooled plasma. Starting from human Cohn fraction IV, we first enriched an Hp-rich fraction using tangential flow filtration (TFF), which was then used as the starting material for purification. Hp was subsequently purified from this fraction using hydrophobic interaction chromatography (HIC) to homogeneity. The final Hp purity reached 98% by SDS-PAGE densitometry and 87% by trypsin digest LC-MS/MS analysis. The purified Hp was further characterized to determine its molecular weight, secondary structure, Hb-binding capacity, and binding kinetics using biophysical techniques including MALDI-TOF, circular dichroism, size exclusion HPLC, and UV-visible stopped-flow spectroscopy. More importantly, the simplicity and efficiency of the TFF-HIC workflow demonstrated strong potential for scalable Hp production.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88555"},"PeriodicalIF":2.8,"publicationDate":"2026-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148900125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

DNA concentration in harvest affects retrovirus clearance capabilities of protein A chromatography. 收获的DNA浓度影响蛋白A层析对逆转录病毒的清除能力。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-09-02 DOI: 10.1002/btpr.88547

Lukas Döring, Sven Schubert, Nora Roos, Johannes Winderl, Annika Müller, Anna Ratermann, Matthias W Kron, Jürgen Hubbuch

{"title":"DNA concentration in harvest affects retrovirus clearance capabilities of protein A chromatography.","authors":"Lukas Döring, Sven Schubert, Nora Roos, Johannes Winderl, Annika Müller, Anna Ratermann, Matthias W Kron, Jürgen Hubbuch","doi":"10.1002/btpr.88547","DOIUrl":"https://doi.org/10.1002/btpr.88547","url":null,"abstract":"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.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88547"},"PeriodicalIF":2.8,"publicationDate":"2026-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148879142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Impact of single-use bioreactor bag materials on Spodoptera frugiperda cell expansion for recombinant adeno-associated virus manufacturing. 一次性生物反应器袋材料对重组腺相关病毒生产夜蛾细胞扩增的影响。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-31 DOI: 10.1002/btpr.88554

Min Tae Park, Seyed Pouria Motevalian

{"title":"Impact of single-use bioreactor bag materials on Spodoptera frugiperda cell expansion for recombinant adeno-associated virus manufacturing.","authors":"Min Tae Park, Seyed Pouria Motevalian","doi":"10.1002/btpr.88554","DOIUrl":"https://doi.org/10.1002/btpr.88554","url":null,"abstract":"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.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88554"},"PeriodicalIF":2.8,"publicationDate":"2026-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148863516","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bone marrow-like viscosity affects imatinib resistance in chronic myeloid leukemia. 骨髓样黏度影响慢性髓性白血病患者对伊马替尼的耐药性。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-22 DOI: 10.1002/btpr.88553

Tianjiao Zeng, Huajian Chen, Man Wang, Toru Yoshitomi, Naoki Kawazoe, Yingnan Yang, Guoping Chen

{"title":"Bone marrow-like viscosity affects imatinib resistance in chronic myeloid leukemia.","authors":"Tianjiao Zeng, Huajian Chen, Man Wang, Toru Yoshitomi, Naoki Kawazoe, Yingnan Yang, Guoping Chen","doi":"10.1002/btpr.88553","DOIUrl":"https://doi.org/10.1002/btpr.88553","url":null,"abstract":"Chronic myeloid leukemia (CML), one type of blood cancer, is a clonal myeloproliferative disorder of hematopoietic stem cells that originated from bone marrow (BM). Imatinib (IM) resistance in CML treatment has been frequently reported, which mainly results from the complex microenvironment of BM. BM is a viscous tissue where cells are mechanically responsive to both the stiffness of matrices and the viscosity of surrounding fluid. Although the effect of stiffness on CML drug resistance has been revealed, the role of BM viscosity on IM resistance in CML is unclear. In this study, a BM-like viscous culture medium was prepared to investigate the effect of microenvironmental viscosity on IM resistance in CML cells. The culture media with viscosity ranging from 47.6 to 634.9 cP were prepared to establish viscosity-controlled culture media covering the reported physiological range of bone marrow and extending into an exploratory higher-viscosity condition. CML cells were suspended and cultured in the viscous culture media added or without addition of IM for 48 or 72 h. Cell viability decreased with IM concentration but increased with microenvironmental viscosity. The results suggested that increased microenvironmental viscosity reduced the sensitivity of CML cells to IM. The possible mechanism might involve the upregulated expression of IM resistance-related genes, reduced apoptosis rates, and altered expression patterns of the BCR::ABL1 gene.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88553"},"PeriodicalIF":2.8,"publicationDate":"2026-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148787917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Lipid droplet profiling during neutrophil differentiation by stimulated Raman scattering microscopy. 用受激拉曼散射显微镜观察中性粒细胞分化过程中的脂滴谱。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-19 DOI: 10.1002/btpr.88552

Ting-Jung Sung, Bin Dong, Jingqiao Shen, Yan Tan, Chi Zhang, Xiaoping Bao

{"title":"Lipid droplet profiling during neutrophil differentiation by stimulated Raman scattering microscopy.","authors":"Ting-Jung Sung, Bin Dong, Jingqiao Shen, Yan Tan, Chi Zhang, Xiaoping Bao","doi":"10.1002/btpr.88552","DOIUrl":"10.1002/btpr.88552","url":null,"abstract":"Lipid droplets (LDs) are dynamic organelles that serve as metabolic hubs and emerging regulators of immune cell fate. Although LDs have been implicated in immune regulation, how LD metabolism is remodeled during neutrophil differentiation and how stage-specific LD dynamics shape mature neutrophil function remain poorly defined. Here, we profiled LD dynamics during neutrophil development and evaluated their role in innate immune function. Using stimulated Raman scattering (SRS) microscopy, we performed label-free, quantitative mapping of LD accumulation in two complementary differentiation systems: murine Hoxb8 myeloid progenitors and human pluripotent stem cell (hPSC)-derived neutrophils. To define metabolic requirements, we pharmacologically modulated LD biosynthesis and catabolism throughout neutrophil differentiation. Perturbing LD metabolism did not impair differentiation efficiency or lineage commitment, but significantly altered the functional output of mature neutrophils. In the murine system, inhibition of adipose triglyceride lipase (ATGL)-mediated LD breakdown enhanced reactive oxygen species (ROS) production and increased anti-tumor cytotoxicity against GL261 glioma cells. In the hPSC model, ATGL inhibition during the myeloid progenitor-to-neutrophil transition selectively increased intracellular LD accumulation without compromising neutrophil yield or purity. This metabolic rewiring also elevated ROS production in hPSC-derived neutrophils, although cytotoxic enhancement against U87MG glioblastoma cells was less pronounced than in the murine system. Collectively, these findings define a stage-specific LD metabolic landscape during neutrophil development and highlight targeted LD modulation as a potential strategy to enhance the functional potency of therapeutic neutrophils.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88552"},"PeriodicalIF":2.8,"publicationDate":"2026-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13548433/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148787836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Multimodal chromatography solutions for the removal of light chain impurity from a fab therapeutic. 从fab治疗剂中去除轻链杂质的多模态色谱溶液。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-17 DOI: 10.1002/btpr.88551

Dongyoun Jang, Nathan Nicholes, Steven M Cramer

{"title":"Multimodal chromatography solutions for the removal of light chain impurity from a fab therapeutic.","authors":"Dongyoun Jang, Nathan Nicholes, Steven M Cramer","doi":"10.1002/btpr.88551","DOIUrl":"https://doi.org/10.1002/btpr.88551","url":null,"abstract":"The production of Fab therapeutics is frequently challenged by an imbalanced expression of light chains (LC) and heavy chains (HC), often resulting in LC-associated impurities. In this work, we evaluated multimodal chromatography as an alternative to affinity chromatography for the removal of an LC impurity. A panel of multimodal CEX and multimodal AEX resins was screened using linear salt and pH gradients to assess Fab monomer and LC impurity selectivities. Product and impurity characterization was carried out using analytical size-exclusion chromatography. Screening data provided insight into the relative contributions of hydrophobic and electrostatic interactions in the Fab and LC impurity selectivities across different multimodal ligands. A previously developed separability scoring metric (Fractional Peak Overlap) was employed to identify Eshmuno HCX and CMM HyperCel as the top resin candidates from the screening data. Finally, bench-scale refinement was carried out to develop an Eshmuno HCX bind-elute step with a 95%-100% recovery and 95%-98% purity of Fab monomer and a CMM HyperCel bind-elute step with a 98%-99% recovery and 98% purity of Fab monomer. This work demonstrates the efficacy of this PD workflow and the utility of multimodal chromatography for removal of LC impurities from Fabs.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88551"},"PeriodicalIF":2.8,"publicationDate":"2026-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148787894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synergistic viral inactivation platform: Rapid enveloped virus inactivation at mildly acidic pH using low non-ionic detergent in Protein A eluates. 协同病毒灭活平台：在温和的酸性pH下使用蛋白A洗脱液中的低非离子洗涤剂快速灭活包膜病毒。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-16 DOI: 10.1002/btpr.88549

June X Zou, Janice H Lee, Shengjiang Shawn Liu, Hendri Tjandra

{"title":"Synergistic viral inactivation platform: Rapid enveloped virus inactivation at mildly acidic pH using low non-ionic detergent in Protein A eluates.","authors":"June X Zou, Janice H Lee, Shengjiang Shawn Liu, Hendri Tjandra","doi":"10.1002/btpr.88549","DOIUrl":"https://doi.org/10.1002/btpr.88549","url":null,"abstract":"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.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88549"},"PeriodicalIF":2.8,"publicationDate":"2026-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148758672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An empirical single-breakthrough curve workflow for rapid early-phase development of continuous capture chromatography for monoclonal antibodies and mAb-derived proteins. 单克隆抗体和单克隆抗体衍生蛋白连续捕获色谱快速早期开发的经验单突破曲线工作流程。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-06 DOI: 10.1002/btpr.88544

Jay Gandhi, James Hilton, Kyo Izumida, Itaru Takeshita, Braden Chiulli, Thomas Higgins, Sami El Hamwi, Janine De Leon, Parinita Bhongale, Evan Graessle, Olga Paley, Mike Dolan, Sheldon Oppenheim, George Parks, Terrence Dobrowsky, Christina Alves

{"title":"An empirical single-breakthrough curve workflow for rapid early-phase development of continuous capture chromatography for monoclonal antibodies and mAb-derived proteins.","authors":"Jay Gandhi, James Hilton, Kyo Izumida, Itaru Takeshita, Braden Chiulli, Thomas Higgins, Sami El Hamwi, Janine De Leon, Parinita Bhongale, Evan Graessle, Olga Paley, Mike Dolan, Sheldon Oppenheim, George Parks, Terrence Dobrowsky, Christina Alves","doi":"10.1002/btpr.88544","DOIUrl":"https://doi.org/10.1002/btpr.88544","url":null,"abstract":"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.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88544"},"PeriodicalIF":2.8,"publicationDate":"2026-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148683221","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

NS0 and CHO cell lines generate distinct quality profiles for non-originator NISTmAbs NS0和CHO细胞系对非起始nistmab产生不同的质量谱。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-05-13 DOI: 10.1002/btpr.88520

Xin Bush, Nicole Azer, JungHyun Kim, David N. Powers, Erica J. Fratz-Berilla

{"title":"NS0 and CHO cell lines generate distinct quality profiles for non-originator NISTmAbs","authors":"Xin Bush, Nicole Azer, JungHyun Kim, David N. Powers, Erica J. Fratz-Berilla","doi":"10.1002/btpr.88520","DOIUrl":"10.1002/btpr.88520","url":null,"abstract":"The selection of host cell lines and culture media plays a central role in influencing the quality attributes of monoclonal antibodies, particularly in biosimilar development and process comparability. This study used standardized upstream conditions to evaluate the impact of host cell line selection on product quality attributes of a non-originator version of the NISTmAb by comparing results from antibodies made in murine myeloma NS0 or Chinese Hamster Ovary (CHO) cells. NISTCHO and NS0 clones achieved comparable peak density and productivity when cultured identically, demonstrating the critical role of media formulation for NISTCHO performance. Distinct cell line-specific differences were noted in aggregation, fragmentation, charge variant distribution, and glycosylation profiles. Non-originator NISTmAb derived from NS0 cells exhibited glycoforms that were nearly 100% fucosylated with higher relative levels of sialylation and galactosylation. The NS0-derived material also displayed elevated levels of acidic charge variant species, decreased main charge variant species, and increased aggregation. Conversely, non-originator NISTmAb derived from NISTCHO exhibited an average of 15–20% relative afucosylation, resulting in an increased binding affinity to FcγRIIIa, which can be of critical importance to quality due to potential enhancement of ADCC potency. These results highlight the intrinsic biology of the host cell line and culture media composition as influential factors for microheterogeneity in post-translational modifications and structural characteristics. As of December 2025, 14 (17.3%) of FDA-approved biosimilars are manufactured using a different cell type than that of the originator product, necessitating comparisons among cell substrates and their effect on product quality of interest for comparative analytical assessment.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147925828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An efficient and robust strategy for developing cell lines through targeted integration 通过靶向整合开发细胞系的有效而稳健的策略。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-08-05 Epub Date: 2026-04-02 DOI: 10.1002/btpr.88509

Duncan McVey, Charu Garg, Chaojie Wang, Aashir Awan, Rui Zheng, Jennifer Zhang, Steve Sturner, Erik Langsdorf, Le Yu, Krishnamurthy Shankarling, Giovanni Rizzi, Kitty Agarwal, Gabi Tremml, Anurag Khetan

{"title":"An efficient and robust strategy for developing cell lines through targeted integration","authors":"Duncan McVey, Charu Garg, Chaojie Wang, Aashir Awan, Rui Zheng, Jennifer Zhang, Steve Sturner, Erik Langsdorf, Le Yu, Krishnamurthy Shankarling, Giovanni Rizzi, Kitty Agarwal, Gabi Tremml, Anurag Khetan","doi":"10.1002/btpr.88509","DOIUrl":"10.1002/btpr.88509","url":null,"abstract":"Reducing the time needed and enhancing predictability of outcomes for cell line development is an ongoing focus of improvement. Targeted integration (TI) where the expression cassettes for a biologic are inserted at a predefined locus, a landing pad, is one of the strategies that is used for this purpose. To be commercially viable the landing pad host cell must support high titers and the transgenes in the landing pad must be highly transcribed. To make such a host TI cell line takes a great deal of time and effort with little guarantee of success. Here we describe a strategy where we converted a stable high-expressing mAb cell line and generated multiple TI host cell lines that are commercially relevant. We routinely generate 6 g/L mAb clones using our platform fit process and 8 to >10 g/L with process optimization that are phenotypically and genetically stable. We also created a duo-landing pad where both landing pads reside in the same locus. This gives the opportunity to simplify the construction of differentially expressed genes since both landing pads are expected to perform equally.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147607502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0