Protein expression and purification最新文献_第3页

Expression and purification of functional recombinant Cullin1-Rbx1/2 in E. coli 功能性重组Cullin1-Rbx1/2在大肠杆菌中的表达与纯化

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-28 DOI: 10.1016/j.pep.2025.106750

Fangyu Zhao , Chuntong Li , Xu Li , Luyu Shi , Yingyue Zhang , Han Wang , Shuzhe Sun , Lu-Jun Liang

{"title":"Expression and purification of functional recombinant Cullin1-Rbx1/2 in E. coli","authors":"Fangyu Zhao , Chuntong Li , Xu Li , Luyu Shi , Yingyue Zhang , Han Wang , Shuzhe Sun , Lu-Jun Liang","doi":"10.1016/j.pep.2025.106750","DOIUrl":"10.1016/j.pep.2025.106750","url":null,"abstract":"<div><div>Protein ubiquitination is a crucial post-translational modification in eukaryotes that is mediated by E1-E2-E3 enzymatic cascades and regulates a wide range of cellular processes. Cullin-RING E3 ligases (CRLs) represent the largest family of RING-type E3 ligases and play pivotal roles in these processes. Generating full-length, active Cullin1-Rbx1 (CRL1) is essential for biochemical and biophysical investigations. In this study, we developed an efficient strategy to produce functional CRL1 complexes from <em>E. coli</em>, by fusing an MsyB protein to N-terminus of Cullin1 to improve its solubility. The recombinant CRL1 demonstrated full functionality, successfully assembling with substrate receptor Skp1-Skp2-Cks1 and mediating the polyubiquitination of Ub-p27-degron substrate. Using recombinant CRL1, we found that phosphorylation at Ser65 inhibited the CRL1-UBE2R1 mediated ubiquitin chain elongation on Ub-p27-degron. Furthermore, using the same strategy, we successfully generated Cullin1-Rbx1 mutants and Cullin1-Rbx2 complexes, thereby expanding the applicability of our method. Collectively, this work establishes a rapid and cost-effective platform for the production of CRL1 complexes, facilitating structural and functional studies.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106750"},"PeriodicalIF":1.4,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144177701","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

A simple protocol for controlling protein deuteration for neutron scattering 控制中子散射中蛋白质氘化的简单规程

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-28 DOI: 10.1016/j.pep.2025.106749

Satoru Fujiwara , Motoyasu Adachi , Yasunobu Sugimoto

{"title":"A simple protocol for controlling protein deuteration for neutron scattering","authors":"Satoru Fujiwara , Motoyasu Adachi , Yasunobu Sugimoto","doi":"10.1016/j.pep.2025.106749","DOIUrl":"10.1016/j.pep.2025.106749","url":null,"abstract":"<div><div>Protein deuteration is a crucial technique in biological studies using neutron scattering/diffraction. In neutron crystallography, the use of deuterated proteins not only reduces the background due to hydrogen atoms but also prevents the cancellation of the scattering length density due to the negative density of hydrogen atoms. In small-angle neutron scattering, it allows the structures of individual components of a protein complex to be studied. Furthermore, in quasielastic neutron scattering, it allows one to measure the dynamics of specific components (or regions) in protein complexes (or a protein). Protocols for protein deuteration employ the expression system in <em>E. coli</em> cultured either in minimal media containing, for example, deuterated glycerol or in rich media containing deuterated algal hydrolysate (algal peptone). While the protocols using the minimal media are the mainstream, the protocols using the rich media allow easy control of deuteration. Here, we optimize the procedure for preparing algal peptone, and describe a protocol for protein deuteration using algal peptone. It is shown that ∼96 % deuteration of the test protein, α-synuclein (αSyn), can be routinely achieved using this protocol. The similarity of the structures of deuterated and hydrogenated αSyn was verified by the small-angle X-ray scattering measurements. Furthermore, the degrees of deuteration can be controlled simply by mixing the hydrogenated and deuterated peptone in the media, and hydrogen labeling of specific amino-acid residues is possible by adding hydrogenated amino acids to the media. The protocol described here is thus useful as a complementary method to those using the minimal media.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106749"},"PeriodicalIF":1.4,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144177702","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

Optimization of CYP27A1 recombinant protein expression CYP27A1重组蛋白表达的优化。

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-26 DOI: 10.1016/j.pep.2025.106748

Johanna E. Papa, Lindsay R. Vaughn, Jackson L. Bartholomew-Schoch, Emma K. Stone, Megen A. Culpepper, Michael J. Reddish

{"title":"Optimization of CYP27A1 recombinant protein expression","authors":"Johanna E. Papa, Lindsay R. Vaughn, Jackson L. Bartholomew-Schoch, Emma K. Stone, Megen A. Culpepper, Michael J. Reddish","doi":"10.1016/j.pep.2025.106748","DOIUrl":"10.1016/j.pep.2025.106748","url":null,"abstract":"<div><div>Human mitochondrial cytochrome P450 27A1 is a monooxygenase enzyme that oxidizes bile acids and other sterol derivatives. The enzyme plays an important role in sterol metabolism and is a potential target for clinical therapies related to metabolic conditions and certain cancers. To support the development of such therapies, detailed structural and functional studies of the enzyme should be pursued. Producing large quantities of purified, recombinant enzyme would enable these studies. Recombinant production of human cytochrome P450 27A1 in <em>E. coli</em> is challenging due to the enzyme being membrane associated. This work explores the optimization of human cytochrome P450 27A1 expression in <em>E. coli</em> by systematically testing the effects of cell strain, expression temperature, concentrations of induction reagents, and expression times. Western blot analysis is used to investigate the effects of variable changes prior to purification. <em>E. coli</em> cell strain (switching to C41(DE3)) appears to have the largest positive effect on overall yield. Increasing δ-aminolevulinic acid concentration (induces heme synthesis) also leads to significantly increased yields. Decreasing expression time decreases the amount of higher order cytochrome P450 aggregates that are formed. The combination of these changes is a more robust expression protocol with three major advantages: decreased expression time, lower aggregate to monomer ratios, and increased overall yield.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106748"},"PeriodicalIF":1.4,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174687","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

Development of magnetic particle-based chemiluminescence immunoassay for quantitative determination of hepatitis C virus core antigen 磁颗粒化学发光免疫法定量测定丙型肝炎病毒核心抗原的建立

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-22 DOI: 10.1016/j.pep.2025.106747

Ye Xu , Yating Liu , Jianrong Zhu , Chufan Tan , Ting Wan , Songhui Zhou , Mengrong Cheng , Jiao Zheng , Rushi Liu

{"title":"Development of magnetic particle-based chemiluminescence immunoassay for quantitative determination of hepatitis C virus core antigen","authors":"Ye Xu , Yating Liu , Jianrong Zhu , Chufan Tan , Ting Wan , Songhui Zhou , Mengrong Cheng , Jiao Zheng , Rushi Liu","doi":"10.1016/j.pep.2025.106747","DOIUrl":"10.1016/j.pep.2025.106747","url":null,"abstract":"<div><h3>Objectives</h3><div>Development of an immunoassay for quantitation detecting the highly conserved hepatitis C core antigen (HCVcAg).</div></div><div><h3>Methods</h3><div>Anti-HCVcAg monoclonal antibodies (mAbs) were developed and evaluated using enzyme-linked immunosorbent assay (ELISA), and the chemiluminescence enzyme immunoassay (CLEIA) was also proposed. The serum HCVcAg was measured to evaluate and analyze the performance of CLEIA and analysis of the mAb pair was evaluated by sequencing of the antibody variable region.</div></div><div><h3>Results</h3><div>The prokaryotically expressed HCVcAg was purifed and three HCVcAg mAbs against HCVcAg with excellent detection performance were obtained after immunizing BALB/c mice. MAb 4D11 and 28B10 were selected as respective capture and detection antibodies for HCVcAg measurement by CLEIA (detection range, 0.1–256 μg/L). The results for CLEIA and fluorescent PCR used in hospitals demonstrated excellent consistency (<em>K</em> = 0.801, <em>P</em> < 0.01). The variable region genes of the heavy chain (VH) and light chain (VL) for the mAb pair were successfully sequenced.</div></div><div><h3>Conclusion</h3><div>The developed mAbs and CLEIA are expected to become a rapid and economical clinical diagnostic tool for HCVcAg detection.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106747"},"PeriodicalIF":1.4,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144138685","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

High-yield production of recombinant CLCF1 protein fused with human serum albumin in animal cells and toxicity evaluation in rodents 重组CLCF1蛋白与人血清白蛋白融合在动物细胞中的高产生产及其对啮齿动物的毒性评价。

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-18 DOI: 10.1016/j.pep.2025.106740

Jeong Min Lee , Jae Sook Kang , Yong Ryoul Yang , Ji Hoon Park

{"title":"High-yield production of recombinant CLCF1 protein fused with human serum albumin in animal cells and toxicity evaluation in rodents","authors":"Jeong Min Lee , Jae Sook Kang , Yong Ryoul Yang , Ji Hoon Park","doi":"10.1016/j.pep.2025.106740","DOIUrl":"10.1016/j.pep.2025.106740","url":null,"abstract":"<div><div>Recombinant protein based biopharmaceutics have been developed as therapeutics of various diseases, especially cancer, diabetes, infectious diseases, and autoimmune diseases. In this study, we conducted a study for the development of biopharmaceuticals based on the CLCF1 protein. First, we established strategies for producing recombinant human CLCF1 protein by transient gene expression in ExpiCHO-S™ Cells and Expi293F™ Cells. For the secretion of CLCF1 protein, we established strategies that human CRLF1 or sCNTFR with CLCF1 protein were co-expressed. As a result, the CLCF1 protein formed a complex with CRLF1 or sCNTFR, which was successfully secreted. Furthermore, the productivity of CLCF1 protein was significantly increased. The ratio of co-transfected plasmids, temperature, CO<sub>2</sub> level and time of harvest were explored, so that the productivity of CLCF1 was remarkably increased 7-fold from 3 mg/L to 22 mg/L. Next, we generated recombinant CLCF1 fusion protein with HSA (Albumin CLCF1) considering the improvement of pharmacokinetic properties and the proven production method in GMP facilities. We evaluated the biological activity of various CLCF1 proteins. In consideration of protein productivity, physical properties, and efficacy, we conducted a single intravenous administration of 4 types of proteins in Sprague-Dawley rats to evaluate the short-term toxicity. As a result, no toxicity related CLCF1 proteins was observed based on the behavior sign observation and body weight changes. In conclusion, we successfully established the strategies of production and characterization of biologically active recombinant CLCF1 proteins in mammalian cells as potential biotherapeutics.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106740"},"PeriodicalIF":1.4,"publicationDate":"2025-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144111266","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 characterization of recombinant epsilon toxin variants of Clostridium perfringens type D in Pichia pastoris D型产气荚膜梭菌Epsilon毒素在毕赤酵母中的表达及特性研究。

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-16 DOI: 10.1016/j.pep.2025.106739

Mauro Joaquin Manfredi, Alejandra Edith Larsen, Eduardo Carlos Mortola, Guillermo Hernán Sguazza

{"title":"Expression and characterization of recombinant epsilon toxin variants of Clostridium perfringens type D in Pichia pastoris","authors":"Mauro Joaquin Manfredi, Alejandra Edith Larsen, Eduardo Carlos Mortola, Guillermo Hernán Sguazza","doi":"10.1016/j.pep.2025.106739","DOIUrl":"10.1016/j.pep.2025.106739","url":null,"abstract":"<div><div>Epsilon toxin (ETX) is the most potent clostridial toxin after tetanus and botulinum neurotoxins. ETX belongs to the heptameric β-pore-forming proteins produced by <em>Clostridium perfringens</em> types B and D, which cause fatal enterotoxemia and significant economic losses in livestock. ETX is synthesized as a protoxin and activated by proteolytic enzymes. Due to limitations in diagnosis and treatment, prevention through vaccination and good management practices is essential. Several commercial vaccines exist for <em>C. perfringens</em> enterotoxemia prevention. However, industrial production is costly and elicits variable immune responses. This has led to the study of recombinant subunit vaccines of ETX with lower toxicity and improved efficacy. The purpose of this study was to produce two versions of the recombinant toxin: truncated epsilon (<em>rETX</em>) and mutated epsilon (<em>rETX</em><sup><em>H106P</em></sup>). Both were cloned and expressed in a <em>Pichia pastoris</em> expression system, followed by small-scale cultivation and a cytotoxicity assay. The assays confirmed that both recombinant polypeptides were non-toxic. Large-scale cultivation conditions were subsequently adjusted to facilitate the evaluation of kinetic parameters and potential expression, focusing on <em>rETX</em><sup><em>H106P</em></sup>. This work demonstrated that <em>rETX</em><sup><em>H106P</em></sup> could potentially be used in a diagnostic kit or as a subunit vaccine candidate against enterotoxemia and other diseases caused by ETX.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106739"},"PeriodicalIF":1.4,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144094689","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

Modular cell line for scalable and rapid in vitro evaluation of chimeric antigen receptors 用于嵌合抗原受体的可扩展和快速体外评价的模块化细胞系

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-13 DOI: 10.1016/j.pep.2025.106738

Simon Dubovik , Dmitri Dormeshkin , Alexandr Migas

{"title":"Modular cell line for scalable and rapid in vitro evaluation of chimeric antigen receptors","authors":"Simon Dubovik , Dmitri Dormeshkin , Alexandr Migas","doi":"10.1016/j.pep.2025.106738","DOIUrl":"10.1016/j.pep.2025.106738","url":null,"abstract":"<div><div>The development of novel Chimeric Antigen Receptor T-cell (CAR-T-cell) products relies on continuous <em>in vitro</em> testing of new antigen/antibody pairs, as well as the modification of the CAR structure and composition. Once Tumor-Associated Antigens (TAA) have been chosen and proven to be specific, and before the first <em>in vivo</em> studies, these CAR compositions must be extensively validated <em>in vitro</em>. Conventional <em>in vitro</em> tests rely on TAA-expressing immortalized cell lines or immortalized cell lines derivatives engineered to stably express the antigen of interest, making this type of model not fully scalable when it comes to multiple TAA/antibody pairs. Therefore, there is a need for a representative, scalable model for preclinical assessment. Here, we introduce a novel model cell line-based approach that can be easily adapted for different experiments by anchoring ALFA-tagged proteins on the cell surface using an anti-ALFA-tag membrane single-domain antibody. This modular multi-component system has the potential to reduce the novel CAR <em>in vitro</em> evaluation procedures from months to weeks.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106738"},"PeriodicalIF":1.4,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144070793","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

Optimization of expression and renaturation methods for the production of a recombinant fibrinolytic protease showing in vivo antithrombotic activity 优化表达和再生方法的生产重组纤溶蛋白酶显示体内抗血栓活性

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-13 DOI: 10.1016/j.pep.2025.106737

Nitisha Boro , Anushree Roy , Ashis K. Mukherjee

{"title":"Optimization of expression and renaturation methods for the production of a recombinant fibrinolytic protease showing in vivo antithrombotic activity","authors":"Nitisha Boro , Anushree Roy , Ashis K. Mukherjee","doi":"10.1016/j.pep.2025.106737","DOIUrl":"10.1016/j.pep.2025.106737","url":null,"abstract":"<div><div>Bacterial fibrinolytic enzymes are promising in treating thrombosis-associated cardiovascular disease. The recombinant fibrinolytic enzymes exhibiting enhanced specificity and improved pharmacokinetics, being less immunogenic and easy to produce, can be advantageous over wild-type enzymes. However, efficient expression and refolding of recombinant enzymes is a significant challenge; therefore, three different analytical methods were compared in this study for the efficient expression and refolding of a recombinant fibrinolytic protease. The gene sequence encoding for fibrinolytic enzyme derived from <em>Bacillus subtilis</em> was codon-optimized according to <em>Escherichia coli</em> codon preference, and this gene was synthetically cloned into the pET26b(+) vector. Alignment of amino acid sequence of this protease gene revealed high sequence similarity with other species of the genus <em>Bacillus</em>. 24 h induced expression at 37 °C and dialysis for renaturation was the most suitable process for expression (enzyme yield) and refolding or renaturation of a ∼40 kDa recombinant α-fibrinogenase enzyme produced in the <em>E. coli</em> (DE3) strain. The recombinant protein demonstrated <em>in vitro</em> fibrinolytic, anticoagulant, thrombin-inhibition, and thrombolytic activities but did not show fibrinogenolytic or <em>in vitro</em> cytotoxicity activity. At a dose of 4 mg/kg, it was found to be non-toxic to Wistar strain albino rats post 72 h of injection but demonstrated dose-dependent <em>in vivo</em> anticoagulant activity.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"233 ","pages":"Article 106737"},"PeriodicalIF":1.4,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144070790","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

A new combination approach to extracellular production of 5-aminolevulinic acid for purification and application in alleviating cadmium-induced oxidative stress in maize 细胞外合成5-氨基乙酰丙酸的新方法及其在镉诱导玉米氧化应激中的应用

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-09 DOI: 10.1016/j.pep.2025.106736

Di Zhang, Jinjing Li, Xiaofeng Chen, Shuncheng Zhang, Baokang Wu, Jun Fan

{"title":"A new combination approach to extracellular production of 5-aminolevulinic acid for purification and application in alleviating cadmium-induced oxidative stress in maize","authors":"Di Zhang, Jinjing Li, Xiaofeng Chen, Shuncheng Zhang, Baokang Wu, Jun Fan","doi":"10.1016/j.pep.2025.106736","DOIUrl":"10.1016/j.pep.2025.106736","url":null,"abstract":"<div><div>5-Aminolevulinic acid (ALA) is widely applied in agriculture, animal husbandry, medicine, and often manufactured in <em>Escherichia coli</em> for overexpressing ALA synthase (ALAS) from α-proteobacteria. For enhancing extracellular ALA production, several approaches have been exploited. Here, we developed and identified a new combination strategy to increase ALA production in <em>E. coli</em>, including selection of the negatively-charged peptide tag as a <em>C</em>-terminal fusion partner for increasing soluble production of the ALAS codon variant from <em>Rhodobacter sphaeroides</em>, mutation of certain residues to increase the ALAS variant activity, optimization of the signal sequences to facilitate ALA secretion, down-regulation of the <em>hemB</em> to inhibit ALA transformation in one plasmid expression system, and supply of 4 mM dithiothreitol to the culture to increase cells tolerant to the oxidative stress. Under the specified cultural conditions, ALA yield was up to 3.2 g/L in flash flasks. Compared with the added cadmium-induced stress, simultaneous supply of purified ALA improved maize seedlings growth, decreased contents of malondialdehyde and hydrogen peroxide, and increased peroxidase activity, contents of chlorophylls and proline.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"232 ","pages":"Article 106736"},"PeriodicalIF":1.4,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143942435","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

Navigating the Frontier: Advances in monoclonal antibody purity control 导航前沿：单克隆抗体纯度控制的进展

IF 1.4 4区生物学

Protein expression and purification Pub Date : 2025-05-07 DOI: 10.1016/j.pep.2025.106725

Xiaoyue Song , Huairu Tian , Rongxian Jing , Kunmei Liu , Kailin Xu , Le Guo , Guolin Zhang

{"title":"Navigating the Frontier: Advances in monoclonal antibody purity control","authors":"Xiaoyue Song , Huairu Tian , Rongxian Jing , Kunmei Liu , Kailin Xu , Le Guo , Guolin Zhang","doi":"10.1016/j.pep.2025.106725","DOIUrl":"10.1016/j.pep.2025.106725","url":null,"abstract":"<div><div>This review explores the evolving landscape of monoclonal antibody (mAb) purity assessment, highlighting the advancements in separation techniques aimed at improving resolution, speed, and precision of impurity detection, identification, and quantification.Traditional methods such as SDS-PAGE, isoelectric focusing electrophoresis and size-exclusion chromatography serve as a foundational tool, while advanced chromatographic techniques, including diverse modes of high-performance liquid chromatography (HPLC), and capillary electrophoresis (CE), offer improved capabilities for analyzing mAb structural integrity and impurities. These techniques separate different constituents in the samples based on their physicochemical properties such as size, charge, isoelectric point, hydrophobicity, and structure. The review discusses the principles and progressions of both traditional and advanced separation techniques and demonstrates their applications in large-scale mAb production and antibody-drug conjugate (ADC) analysis. Innovations in chromatography, such as superficially porous particles and novel stationary phases, address challenges like band broadening and protein adsorption.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"232 ","pages":"Article 106725"},"PeriodicalIF":1.4,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143937480","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