Protein expression and purification最新文献_第6页

Modulation of N-glycosylation in bispecific antibody biosimilars through combined modulators 通过联合调节剂调节双特异性抗体生物类似药的n -糖基化。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-09-26 DOI: 10.1016/j.pep.2025.106825

Xu Shengnan , Wang Pan , Wang Xiaofei, Liu Xiaojing, Li Guozhu, Qin Guohong, Xu Dan

{"title":"Modulation of N-glycosylation in bispecific antibody biosimilars through combined modulators","authors":"Xu Shengnan , Wang Pan , Wang Xiaofei, Liu Xiaojing, Li Guozhu, Qin Guohong, Xu Dan","doi":"10.1016/j.pep.2025.106825","DOIUrl":"10.1016/j.pep.2025.106825","url":null,"abstract":"<div><div>To ensure a high degree of similarity between biosimilars and reference drugs, it is crucial to perform comprehensive characterization and maintain rigorous control over glycosylation processes. Here,we aimed to optimize the glycosylation profile of a biosimilar of a bispecific antibody (BsAb) to closely resemble that of the reference drug through the synergistic use of glycosylation modulators. To identify the strongest modulators and appropriate concentration ranges, we first examined the effects of different concentrations of galactose (Gal) and manganese chloride (MnCl<sub>2</sub>) on the galactosylation rate in Shake Flask, as well as the influence of tris(hydroxymethyl)aminomethane (Tris) on the incorporation of mannose and fucose in 2 L Bioreactor. Importantly, the concurrent use of Tris and galactose did not result in any interaction effects on N-glycan modifications and had no detrimental impact on cell growth, metabolism, antibody charge variants or purity. In conclusion, The concurrent use of 0.75 mM Tris and 8 mM galactose yields a glycosylation profile of Bs-mAb1 that is highly comparable to that of the reference drug, thereby providing an effective strategy for optimizing glycosylation in biosimilars. These findings provide significant insights into the regulation of glycosylation in the production of therapeutic monoclonal antibodies and may contribute to enhancing the consistency and therapeutic performance of biosimilars.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106825"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145186603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Expression and purification of Austropuccinia psidii effector proteins in Escherichia coli psidii效应蛋白在大肠杆菌中的表达与纯化。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-09-16 DOI: 10.1016/j.pep.2025.106815

Jovarn V. Sullivan , Michael J. Currie , Vanessa K. Morris , Ashish Sethi , Santosh Panjikar , Grant R. Smith , Claudia-Nicole Meisrimler , Renwick C.J. Dobson

{"title":"Expression and purification of Austropuccinia psidii effector proteins in Escherichia coli","authors":"Jovarn V. Sullivan , Michael J. Currie , Vanessa K. Morris , Ashish Sethi , Santosh Panjikar , Grant R. Smith , Claudia-Nicole Meisrimler , Renwick C.J. Dobson","doi":"10.1016/j.pep.2025.106815","DOIUrl":"10.1016/j.pep.2025.106815","url":null,"abstract":"<div><div>The plant disease myrtle rust is caused by the fungus <em>Austropuccinia psidii</em>. It has led to functional myrtaceous species extinctions in Australia and is a significant threat to other species globally. During infection, <em>A. psidii</em> secretes effector proteins that manipulate the host plant's defences. Numerous putative effectors are encoded in this pathogen's genome, some being expressed early during urediniospore germination and initial invasion of plant tissues. Four putative effector proteins (AP1260, AP5292, AP10948, and AP143) were found to be differentially expressed in the first 24–48 h of infection, suggesting that they play important roles in the infection process. As in other rust fungi, these effector proteins are small and cysteine-rich, often forming disulfide bonds, and their isolation for biophysical characterisation can be challenging. AlphaFold3 models predict that AP1260, AP5292, AP10948, and AP143 form disulfide bonds, while disorder analysis indicates the presence of intrinsically disordered regions. The four putative <em>A. psidii</em> effector proteins were recombinantly produced using SHuffle <em>Escherichia coli</em> cells with an adapted co-expression vector, ‘ApFunCyDisCo’. Three of the effectors were successfully produced, but were insoluble. The fourth effector, AP1260, was successfully produced in the soluble fraction and purified using a four-step process: immobilised metal affinity chromatography, desalting, anion exchange chromatography, and size exclusion chromatography. Circular dichroism spectroscopy revealed that AP1260 has a mainly random coil character, but also has both β-strand and α-helical content. This first successful production and isolation of an <em>A. psidii</em> protein provides a foundation for future investigation of the molecular mechanisms of <em>A. psidii</em> pathogenicity.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106815"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145086749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Adaptation of induced pluripotent stem cell technology for avian species 诱导多能干细胞技术在鸟类中的适应性研究。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-09-11 DOI: 10.1016/j.pep.2025.106814

Masafumi Katayama , Tomokazu Fukuda

{"title":"Adaptation of induced pluripotent stem cell technology for avian species","authors":"Masafumi Katayama , Tomokazu Fukuda","doi":"10.1016/j.pep.2025.106814","DOIUrl":"10.1016/j.pep.2025.106814","url":null,"abstract":"<div><div>Following the first report of induced pluripotent stem (iPS) cells from mouse, various mammalian-derived iPS cells have been established. In contrast, avian-derived iPS cells or iPS-like cells have been rarely reported. iPS cells can differentiate into various cell types (e.g., neural cells and hepatocytes) and proliferate indefinitely in culture. Unlike embryonic stem cells, iPS cells are generated from somatic cells, eliminating the need for embryos in their generation. Because somatic cells can be obtained from deceased individuals, iPS cell technology can be adapted for use in wild avian species, beyond its application in chickens. Our group previously reported the generation of chicken iPS cells from somatic cells using a modified-octamer-binding transcription factor 3/4 (Oct3/4), SRY-box transcription factor 2 (Sox2), Krüppel-like factor 4 (Klf4), MYC proto-oncogene (c-Myc), Nanog, and lin-28 homolog A (Lin28A). Developmental chicken eggs are a valuable resource for protein production. We may obtain valuable proteins from the chimeric developmental eggs of chickens using genome-edited or transgenic chicken iPS cells. Furthermore, our group established iPS cells derived from endangered avian species (Okinawa rail, Japanese ptarmigan, and Blakiston's fish owl) using modified-Oct3/4, Sox2, Klf4, c-Myc, Nanog, Lin28, and Klf2. Cells differentiated from iPS cells (e.g., neural cells and hepatocytes) can be used for drug testing in veterinary medicine and for evaluating sensitivity to infectious diseases and pollutants. We believe that iPS cell technology can be developed as a powerful tool for the conservation of endangered avian species.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106814"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145058524","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Expression and purification of C14orf119 and generation of its polyclonal antibody C14orf119的表达纯化及其多克隆抗体的制备。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-09-22 DOI: 10.1016/j.pep.2025.106822

Yong-Hong Nie , Qiang Li , Yan-Ji Lu , Tuo Tang , Xian Hong , Xin-Xin Yue , Zhi-Hui Deng , Tao Wang

{"title":"Expression and purification of C14orf119 and generation of its polyclonal antibody","authors":"Yong-Hong Nie , Qiang Li , Yan-Ji Lu , Tuo Tang , Xian Hong , Xin-Xin Yue , Zhi-Hui Deng , Tao Wang","doi":"10.1016/j.pep.2025.106822","DOIUrl":"10.1016/j.pep.2025.106822","url":null,"abstract":"<div><div>C14orf119 is a functionally uncharacterized mitochondrial protein potentially implicated in ischemic stroke pathogenesis. To enable comprehensive biological studies, we developed and validated a specific polyclonal antibody against this target. Our approach involved prokaryotic expression and chromatographic purification of both His- and GST-tagged C14orf119 in <em>Escherichia coli</em>. The purified His-C14orf119 served as immunogen for rabbit polyclonal antibody production, while the purified GST-C14orf119 enabled subsequent affinity purification of target-specific antibody. Rigorous characterization demonstrated the antibody's exclusive specificity for both exogenous and endogenous C14orf119. Notably, the C14orf119 antibody failed to detect control His-tagged proteins, confirming effective removal of anti-His antibodies during the GST-based affinity purification process. This rigorously validated antibody provides a critical molecular tool for functional characterization of C14orf119, paving the way for mechanistic investigations into its pathophysiological roles.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106822"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Secretory expression and fermentation optimization of recombinant porcine epidermal growth factor in Saccharomyces cerevisiae 重组猪表皮生长因子在酿酒酵母中的分泌表达及发酵优化。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-10-08 DOI: 10.1016/j.pep.2025.106827

Jie Liu , Wenjun Xu , Bingbing Xia , Yanfei He , Shiping Huang , Jun Zhao

{"title":"Secretory expression and fermentation optimization of recombinant porcine epidermal growth factor in Saccharomyces cerevisiae","authors":"Jie Liu , Wenjun Xu , Bingbing Xia , Yanfei He , Shiping Huang , Jun Zhao","doi":"10.1016/j.pep.2025.106827","DOIUrl":"10.1016/j.pep.2025.106827","url":null,"abstract":"<div><div>Porcine epidermal growth factor (PoEGF) promotes intestinal epithelial development and enhances immune function in weaned piglets, offering potential as an alternative to in-feed antibiotics. However, large-scale application of recombinant PoEGF (rPoEGF) is hindered by high production costs and complex purification requirements. In this study, we constructed a recombinant <em>Saccharomyces cerevisiae</em> INVSc1 strain capable of expressing and secreting rPoEGF through the integration of an α-factor signal peptide-PoEGF-6 × His expression cassette. Expression was confirmed by SDS-PAGE and Western blot analysis, and bioactivity was validated using a BALB/c 3T3 fibroblast proliferation assay, yielding a maximum mitogenic activity of 10,963 U/mL. To improve production efficiency, a two-stage fermentation strategy was developed and optimized in a 4-L bioreactor. The process consisted of a glucose-based fed-batch growth phase followed by galactose-induced expression under optimized conditions (25 °C, pH 5.0, DO 30 %). This approach enabled high-cell-density cultivation (OD<sub>600</sub> ≈ 71.5) and achieved a final secreted protein yield of 30.2 mg/L. Notably, the culture containing active rPoEGF could be directly applied as a functional feed additive without the need for downstream purification. These results provide a practical, scalable, and cost-effective approach for producing bioactive rPoEGF and support its future application in swine production as a safe, antibiotic-free growth promoter.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106827"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145259007","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Transcriptome profiles for defining avian primordial germ cell development 定义禽原始生殖细胞发育的转录组谱。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-09-11 DOI: 10.1016/j.pep.2025.106812

Kennosuke Ichikawa, Mike J. McGrew

引用次数: 0

Biochemical characterization of histidinol dehydrogenase from the human pathogen Neisseria gonorrhoeae 淋病奈瑟菌组氨酸二醇脱氢酶的生化特性研究。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-09-26 DOI: 10.1016/j.pep.2025.106824

Shuping Fei , Jiatong Zhao , Wenwen Wu , Chaoqun Lian , Wanggang Tang

{"title":"Biochemical characterization of histidinol dehydrogenase from the human pathogen Neisseria gonorrhoeae","authors":"Shuping Fei , Jiatong Zhao , Wenwen Wu , Chaoqun Lian , Wanggang Tang","doi":"10.1016/j.pep.2025.106824","DOIUrl":"10.1016/j.pep.2025.106824","url":null,"abstract":"<div><div>Histidinol dehydrogenase (HisD, E.C. 1.1.1.23), encoded by the <em>hisD</em> gene and catalyzing the final two steps in the <span>l</span>-histidine biosynthesis, has emerged as a promising antibacterial target for several human pathogens, such as <em>Brucella suis</em> and <em>Mycobacterium tuberculosis</em>. Herein, biochemical properties on recombinant HisD from the human pathogen <em>Neisseria gonorrhoeae</em> (<em>Ng</em>HisD) were characterized in detail. SDS-PAGE and size-exclusion chromatography revealed that recombinant <em>Ng</em>HisD is a homodimer (∼105 kDa native size; ∼47 kDa subunit). Kinetic analysis identified this protein as a highly selective NAD<sup>+</sup>-dependent enzyme (specific activity: 17.3 ± 0.4 U mg<sup>−1</sup>), exhibiting minimal activity with NADP<sup>+</sup>. The <em>K</em><sub>M</sub> values for NAD<sup>+</sup> and <span>l</span>-histidinol were 0.9 ± 0.3 mM and 45 ± 6 μM, respectively. Maximal activity occurred at pH 9.5 and 45 °C in the presence of Mn<sup>2+</sup>. Heat-inactivation experiment showed that rapid inactivation began to occur following incubation at 45 °C for 30 min. The metal ions coordinated by the enzyme were not sequestered by 10 mM EDTA but were efficiently chelated by 1,10-phenanthroline. The recombinant <em>Ng</em>HisD lost 85 % activity in the presence of 1 mM 1,10-phenantroline. Our study not only establishes a foundation for investigating the physiological function of <em>Ng</em>HisD <em>in vivo</em> but also uncovers the absence of the canonical histidine biosynthesis pathway in <em>Neisseria cinerea</em>.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106824"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145186531","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Circular dichroism spectroscopy in protein engineering and pharmaceutical development: Applications in structural characterization and quality assessment 圆二色光谱在蛋白质工程和药物开发中的应用：结构表征和质量评价。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-10-08 DOI: 10.1016/j.pep.2025.106826

Taiji Oyama, Satoko Suzuki, Ken-ichi Akao

{"title":"Circular dichroism spectroscopy in protein engineering and pharmaceutical development: Applications in structural characterization and quality assessment","authors":"Taiji Oyama, Satoko Suzuki, Ken-ichi Akao","doi":"10.1016/j.pep.2025.106826","DOIUrl":"10.1016/j.pep.2025.106826","url":null,"abstract":"<div><div>Circular dichroism (CD) spectroscopy is a rapid and versatile method for assessing the higher-order structures of proteins and peptides. Far-UV CD enables quantitative evaluation of secondary structures, while near-UV CD provides sensitive fingerprints of tertiary organization. With advances in recombinant protein expression, CD spectroscopy has become widely used for the characterization of novel generated proteins. This review focuses on the applications of CD spectroscopy in protein engineering and pharmaceutical sciences. Examples include strategies for reliable measurements with limited sample quantities and quality assessments such as lot-to-lot comparisons and biosimilar evaluations. CD spectroscopy also serves as a valuable tool for detecting conformational changes associated with protein–protein and protein–drug interactions, as well as for evaluating proteins and peptides in membrane-mimetic environments. Obtaining reliable CD spectra requires careful selection of buffers, water-miscible solvents, and excipients. Here, we summarize their properties and propose practical criteria for their selection to ensure high-quality CD measurements.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106826"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145259018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Validation of cold shock-based technique for purification of recombinant C. elegans Phosphodiesterase 3 protein expressed in E. coli 冷冲击技术纯化重组秀丽隐杆线虫磷酸二酯酶3蛋白的研究。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-07-25 DOI: 10.1016/j.pep.2025.106769

Swapna G. Naik , Dong Keun Rhee , Steven Hockman , Faiyaz Ahmad Khan , Arun Samidurai , Babak Sabouri , Noel Carter , Vincent C. Manganiello

{"title":"Validation of cold shock-based technique for purification of recombinant C. elegans Phosphodiesterase 3 protein expressed in E. coli","authors":"Swapna G. Naik , Dong Keun Rhee , Steven Hockman , Faiyaz Ahmad Khan , Arun Samidurai , Babak Sabouri , Noel Carter , Vincent C. Manganiello","doi":"10.1016/j.pep.2025.106769","DOIUrl":"10.1016/j.pep.2025.106769","url":null,"abstract":"<div><div>Phosphodiesterase 3 enzymes (PDE3) play important roles in the regulation of adipocyte lipolysis and cardiac contractility by hydrolyzing cAMP and cGMP. This study reports the cloning, expression, and purification of recombinant <em>Caenorhabditis elegans</em> phosphodiesterase 3 (CEPDE3) isoforms, using a cold shock-based technique. The two closely related isoforms of the <em>CEPDE3</em> gene (isoform F and isoform A) were cloned into the pGEX-6P-1 vector and expressed in <em>E. coli</em> as fusion proteins with a glutathione-S transferase (GST) tag at their amino terminus and purified by affinity chromatography using a glutathione Sepharose column. To optimize expression and recovery of soluble CEPDE3 protein from <em>E. coli</em>, we applied a “cold shock” technique at 4 °C following IPTG induction. Our findings suggest improved protein expression using an N-terminal GST tag instead of a C-terminal 6-histidine (6His) tag. Exposure of GST-tagged CEPDE3 to cold shock improved the protein solubility of CEPDE3 isoforms recovered by affinity chromatography. Studying CEPDE3 expression in <em>C. elegans</em> may enable us to understand the structure-function relationship and help crystallize the proteins to identify its catalytic pocket, contributing to the design of more effective small modulators of PDE3 catalytic activity.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106769"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144732922","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Effective and robust separation of half-antibody byproduct in bispecific antibody purification by Sartobind Rapid A Protein A membrane chromatography Sartobind快速A蛋白A膜层析纯化双特异性抗体时半抗体副产物的有效和稳健分离。

IF 1.2 4区生物学

Protein expression and purification Pub Date : 2026-01-01 Epub Date: 2025-09-25 DOI: 10.1016/j.pep.2025.106823

Gaoya Yuan, Meng Qu, Yifeng Li

{"title":"Effective and robust separation of half-antibody byproduct in bispecific antibody purification by Sartobind Rapid A Protein A membrane chromatography","authors":"Gaoya Yuan, Meng Qu, Yifeng Li","doi":"10.1016/j.pep.2025.106823","DOIUrl":"10.1016/j.pep.2025.106823","url":null,"abstract":"<div><div>Half-antibody is a common byproduct associated with the production of asymmetric bispecific antibody (bsAb). We previously demonstrated that Protein A column chromatography can effectively separate this byproduct from the bsAb product, as these two species exhibit different binding valencies (half-antibody and intact bsAb contain one and two Protein A binding sites, respectively). Specifically, half-antibody, which binds weaker, can be selectively removed by an appropriate wash prior to elution of the target bsAb. However, the performance of this wash step is sensitive to the loading density, making the process unrobust to variations in harvest titer. In the current study, we demonstrate that Sartobind Rapid A Protein A membrane effectively reduces half-antibody from approximately 16.5% (in the load) to 0.1% (in the eluate). In addition, using Protein A membrane also improves process robustness by avoiding the impact of loading density variation on performance. Thus, for bsAb purification where half-antibody removal is required, Protein A membrane is a better choice than resin-based Protein A column.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"237 ","pages":"Article 106823"},"PeriodicalIF":1.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182205","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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