Biotechnology and Bioengineering最新文献_第5页

Buffer system improves the removal of host cell protein impurities in monoclonal antibody purification 缓冲液系统可改善单克隆抗体纯化过程中宿主细胞蛋白质杂质的去除效果

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-18 DOI: 10.1002/bit.28844

Dániel Lakatos, Martina Idler, Selina Stibitzky, Jennifer Amann, Jakob Schuschkewitz, Dominik Krayl, Judith Liebau, Jan-Hendrik Grosch, Erik Arango Gutierrez, Simon Kluters

{"title":"Buffer system improves the removal of host cell protein impurities in monoclonal antibody purification","authors":"Dániel Lakatos, Martina Idler, Selina Stibitzky, Jennifer Amann, Jakob Schuschkewitz, Dominik Krayl, Judith Liebau, Jan-Hendrik Grosch, Erik Arango Gutierrez, Simon Kluters","doi":"10.1002/bit.28844","DOIUrl":"10.1002/bit.28844","url":null,"abstract":"Polysorbates (PS) are commonly used as stabilizers of biopharmaceuticals such as monoclonal antibodies (mAbs). However, they are prone to chemical and enzymatic degradation. The latter can be caused by residual host cell proteins (HCPs) in the drug substance. Degradation affects the functionality of the PS surfactant which can lead to formation of particles. An increasing number of publications describe enzymatic PS degradation. Significant efforts have been made to characterize HCP removal during Downstream Processing (DSP) of mAbs and to develop mitigation strategies. Here we describe the use of glycine buffer for acidic elution in Protein A affinity chromatography compared to acetate buffer, which is more commonly used in the biopharmaceutical industry. Increased turbidity was observed during pH re-adjustment after low pH virus inactivation when using glycine buffer. Analytical data suggests that this turbidity is caused by the formation of precipitates which include HCP and DNA impurities. Additionally, as a zwitterion, glycine does not contribute to conductivity; this further enhances HCP removal during anion-exchange flow-through chromatography. Although glycine is well known as a possible elution buffer for Protein A affinity chromatography, its positive impact on HCP removal and PS stability have not yet been described in literature.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 12","pages":"3869-3880"},"PeriodicalIF":3.5,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28844","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142246059","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

3D printed gelatin methacryloyl hydrogels for perfusion culture of human trabecular meshwork cells and glaucoma studies 用于人类小梁网细胞灌注培养和青光眼研究的三维打印明胶甲基丙烯酰水凝胶

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-18 DOI: 10.1002/bit.28849

Bikram Adhikari, Prasanga Barakoti, Mina B. Pantcheva, Melissa D. Krebs

引用次数: 0

Physics-informed neural networks for biopharmaceutical cultivation processes: Consideration of varying process parameter settings 用于生物制药培养过程的物理信息神经网络：考虑不同的工艺参数设置

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-18 DOI: 10.1002/bit.28851

Niklas Adebar, Sabine Arnold, Liliana M. Herrera, Victor N. Emenike, Thomas Wucherpfennig, Jens Smiatek

{"title":"Physics-informed neural networks for biopharmaceutical cultivation processes: Consideration of varying process parameter settings","authors":"Niklas Adebar, Sabine Arnold, Liliana M. Herrera, Victor N. Emenike, Thomas Wucherpfennig, Jens Smiatek","doi":"10.1002/bit.28851","DOIUrl":"https://doi.org/10.1002/bit.28851","url":null,"abstract":"We present a new modeling approach for the study and prediction of important process outcomes of biotechnological cultivation processes under the influence of process parameter variations. Our model is based on physics-informed neural networks (PINNs) in combination with kinetic growth equations. Using Taylor series, multivariate external process parameter variations for important variables such as temperature, seeding cell density and feeding rates can be integrated into the corresponding kinetic rates and the governing growth equations. In addition to previous approaches, PINNs also allow continuous and differentiable functions as predictions for the process outcomes. Accordingly, our results show that PINNs in combination with Taylor-series expansions for kinetic growth equations provide a very high prediction accuracy for important process variables such as cell densities and concentrations as well as a detailed study of individual and combined parameter influences. Furthermore, the proposed approach can also be used to evaluate the outcomes of new parameter variations and combinations, which enables a saving of experiments in combination with a model-driven optimization study of the design space.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"13 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142246058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Culture system for longitudinal monitoring of bone dynamics ex vivo 用于纵向监测体内外骨骼动态的培养系统

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-18 DOI: 10.1002/bit.28848

Esther E.A. Cramer, Kim C.J. Hermsen, Linda M. Kock, Keita Ito, Sandra Hofmann

{"title":"Culture system for longitudinal monitoring of bone dynamics ex vivo","authors":"Esther E.A. Cramer, Kim C.J. Hermsen, Linda M. Kock, Keita Ito, Sandra Hofmann","doi":"10.1002/bit.28848","DOIUrl":"https://doi.org/10.1002/bit.28848","url":null,"abstract":"To quantify and visualize both bone formation and resorption within osteochondral explants cultured ex vivo is challenging with the current analysis techniques. An approach that enables monitoring of bone remodeling dynamics is longitudinal microcomputed tomography (µCT), a non-destructive technique that relies on repeated µCT scanning and subsequent registration of consecutive scans. In this study, a two-compartment culture system suitable for osteochondral explants that allowed for µCT scanning during ex vivo culture was established. Explants were scanned repeatedly in a fixed orientation, which allowed assessment of bone remodeling due to adequate image registration. Using this method, bone formation was found to be restricted to the outer surfaces when cultured statically. To demonstrate that the culture system could capture differences in bone remodeling, explants were cultured statically and under dynamic compression as loading promotes osteogenesis. No quantitative differences between static and dynamic culture were revealed. Still, only in dynamic conditions, bone formation was visualized on trabecular surfaces located within the inner cores, suggesting enhanced bone formation towards the center of the explants upon mechanical loading. Taken together, the ex vivo culture system in combination with longitudinal µCT scanning and subsequent registration of images demonstrated potential for evaluating bone remodeling within explants.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"195 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142246060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Clear insight into complex multimodal resins and impurities to overcome recombinant protein purification challenges: A review 清晰洞察复杂的多模式树脂和杂质，克服重组蛋白纯化难题：综述

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-18 DOI: 10.1002/bit.28846

Maryam Moazami Goodarzi, Reza Jalalirad

{"title":"Clear insight into complex multimodal resins and impurities to overcome recombinant protein purification challenges: A review","authors":"Maryam Moazami Goodarzi, Reza Jalalirad","doi":"10.1002/bit.28846","DOIUrl":"https://doi.org/10.1002/bit.28846","url":null,"abstract":"Increasing attention has been paid to the purity of therapeutic proteins imposing extensive costs and challenges to the downstream processing of biopharmaceuticals. One of the efforts, that has been exerted to overcome such limitations, was developing multimodal or mixed‐mode chromatography (MMC) resins for launching selective, orthogonal, non‐affinity purification platforms. Despite relatively extensive usage of MMC resins, their real potential and fulfillment have not been extensively reviewed yet. In this work, the explanation of practical and key aspects of downstream processing of recombinant proteins with or without MMC resins was debated, as being useful for further purification process development. This review has been written as a step‐by‐step guide to deconvolute both inherent protein purification and MMC complexities. Here, after complete elucidation of the potential of MMC resins, the effects of frequently used additives (mobile phase modifiers) and their possible interactions during the purification process, the critical characteristics of common product‐related impurities (e.g., aggregates, charge variants, fragments), host‐related impurities (e.g., host cell protein and DNA) and process related impurities (e.g., endotoxin, and viruses) with solved or unsolved challenges of traditional and MMC resins have been discussed. Such collective experiences which are reported in this study could be considered as an applied guide for developing successful downstream processing in challenging conditions by providing a clear insight into complex MMC resins and impurities.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"33 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142236280","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Active diagnostic ingredients (ADIs) for PCR: A mini-bioreactor producing dNTPs with silica immobilized R5-kinases 用于 PCR 的活性诊断成分 (ADI)：用硅胶固定 R5 激酶生产 dNTPs 的微型生物反应器

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-18 DOI: 10.1002/bit.28837

Anna R. Bird, Elizabeth A. H. Hall

{"title":"Active diagnostic ingredients (ADIs) for PCR: A mini-bioreactor producing dNTPs with silica immobilized R5-kinases","authors":"Anna R. Bird, Elizabeth A. H. Hall","doi":"10.1002/bit.28837","DOIUrl":"10.1002/bit.28837","url":null,"abstract":"Low availability of routine nucleic acid amplification testing (NAAT) during infection outbreaks, especially in less resourced environments, was highlighted by the Covid pandemic. One of the barriers lies with the supply chain and cost of the active diagnostic ingredients (ADIs) that are the reagents for NAATs. This work explores a novel synthesis method to produce a key NAAT reagent, namely the 2′-deoxynucleoside 5′-triphosphate (dNTPs), via a reusable enzyme bioreactor, that can be integrated into a NAAT workflow. A self-immobilizing R5-silaffin kinase fusion enzyme was designed for immobilization on silica, converting dNMPs to their respective dNTP ADIs for PCR in a R5-kinase mini-bioreactor, designed to be implemented in a reusable device, stable over 2 months, when stored at 4°C. The performance is demonstrated for PCR reactions of the lambda genome and showed successful amplification up to 7.5 kb. In comparison with commercial dNTPs, in Plasmodium malariae NAATs, a high linear correlation was shown between the Ct value and the log(Copy Number), with lower incidence of false positives than with the commercial dNTPs. Overall a pathway to generate deoxynucleotides from monophosphate precursors was demonstrated, and an immobilized enzyme mini-bioreactor investigated as a proof-of-principle for work-flow integration with NAAT in low-resource research and diagnostics labs.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 12","pages":"3834-3847"},"PeriodicalIF":3.5,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28837","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142237017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

3D bioprinting technology and equipment based on microvalve control 基于微阀控制的 3D 生物打印技术和设备

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-17 DOI: 10.1002/bit.28850

Rihui Kang, Jiaxing Wu, Rong Cheng, Meng Li, Luxiao Sang, Hulin Zhang, Shengbo Sang

引用次数: 0

Electrobiofabrication of antibody sensor interfaces within a 3D printed device yield rapid and robust electrochemical measurements of titer and glycan structure 在三维打印设备中电生物制造抗体传感器接口，实现快速、稳健的滴度和聚糖结构电化学测量

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-16 DOI: 10.1002/bit.28839

Chen-Yu Chen, Dana Motabar, Fauziah Rahma Zakaria, Eunkyoung Kim, Benjamin Wu, Gregory F. Payne, William E. Bentley

{"title":"Electrobiofabrication of antibody sensor interfaces within a 3D printed device yield rapid and robust electrochemical measurements of titer and glycan structure","authors":"Chen-Yu Chen, Dana Motabar, Fauziah Rahma Zakaria, Eunkyoung Kim, Benjamin Wu, Gregory F. Payne, William E. Bentley","doi":"10.1002/bit.28839","DOIUrl":"10.1002/bit.28839","url":null,"abstract":"We report the integration of 3D printing, electrobiofabrication, and protein engineering to create a device that enables near real-time analysis of monoclonal antibody (mAb) titer and quality. 3D printing was used to create the macroscale architecture that can control fluidic contact of a sample with multiple electrodes for replicate measurements. An analysis “chip” was configured as a “snap-in” module for connecting to a 3D printed housing containing fluidic and electronic communication systems. Electrobiofabrication was used to functionalize each electrode by the assembly of a hydrogel interface containing biomolecular recognition and capture proteins. Specifically, an electrochemical thiol oxidation is used to assemble a thiolated polyethylene glycol hydrogel, that in turn is covalently coupled to either a cysteine-tagged protein G that binds the antibody's Fc region or a lectin that binds the glycans of target mAb analytes. We first show the design, assembly, and testing of the hardware device. Then, we show the transition of a step-by-step sensing methodology (e.g., mix, incubate, wash, mix, incubate, wash, measure) into the current method where functionalization, antibody capture, and assessment are performed in situ and in parallel channels. Both titer and glycan analyses were found to be linear with antibody concentration (to 0.2 mg/L). We further found the interfaces could be reused with remarkably similar results. Because the interface assembly and use are simple, rapid, and robust, we suggest this assessment methodology will be widely applicable, including for other biomolecular process development and manufacturing environments.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 12","pages":"3754-3767"},"PeriodicalIF":3.5,"publicationDate":"2024-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28839","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142237020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Development of a multigene expression system using 2A peptides in Rhodosporidium toruloides 利用 Toruloides 罗多孢子虫中的 2A 肽开发多基因表达系统

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-16 DOI: 10.1002/bit.28843

Xiao Guo, Zhenzhen Bai, Huimin Zhao, Shuobo Shi

引用次数: 0

Engineering of heterobifunctional biopolymers for tunable binding and precipitation of Strep-Tag proteins and virus-like nanoparticles 设计异多功能生物聚合物，实现 Strep-Tag 蛋白和类病毒纳米粒子的可调结合与沉淀

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-09-15 DOI: 10.1002/bit.28845

James Tang, Matthew Becker, Abraham Lenhoff, Wilfred Chen

{"title":"Engineering of heterobifunctional biopolymers for tunable binding and precipitation of Strep-Tag proteins and virus-like nanoparticles","authors":"James Tang, Matthew Becker, Abraham Lenhoff, Wilfred Chen","doi":"10.1002/bit.28845","DOIUrl":"10.1002/bit.28845","url":null,"abstract":"Affinity precipitation is a powerful separation method in that it combines the binding selectivity of affinity chromatography with precipitation of captured biomolecules via phase separation triggered by small changes in the environment, e.g., pH, ionic strength, temperature, light, etc. Elastin-like polypeptides (ELPs) are thermally responsive biopolymers composed of pentapeptide repeats VPGVG that undergo reversible phase separation, where they aggregate when temperature and/or salt concentration are increased. Here we describe the generation of an ELP fusion to a soluble streptavidin mutant that enables rapid purification of any Strep-tag II fusion protein of interest. This heterobifunctional protein takes advantage of the native tetrameric structure of streptavidin, leading to binding-induced multivalent crosslinking upon protein capture. The efficient biotin-mediated dissociation of the bound Strep-tag II fusion protein from the streptavidin-ELP capturing scaffold allows for mild elution conditions. We also show that this platform is particularly effective in the purification of a virus-like particle (VLP)-like E2 protein nanoparticle, likely because the high valency of the protein particle causes binding-induced crosslinking and precipitation. Considering the importance of VLP for gene therapy applications, we believe this is a particularly exciting advance. We demonstrated this feasibility by the efficient purification of a VLP-like E2 protein nanoparticle as a surrogate.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 12","pages":"3860-3868"},"PeriodicalIF":3.5,"publicationDate":"2024-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142234052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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