Protein expression and purification最新文献_第6页

Development and expression of a factor X-derived recombinant protein for ELISA-based detection of Emicizumab 基于elisa检测Emicizumab的因子x衍生重组蛋白的开发和表达。

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification

Pub Date : 2026-02-01 Epub Date: 2025-10-14 DOI: 10.1016/j.pep.2025.106829

Sina Tavakoli , Shohreh Zare Karizi , Jafar Amani

Emicizumab is a therapeutic antibody used in the management of Hemophilia A. It mimics the function of coagulation Factor VIII by bridging activated Factor IX (FXa) and Factor X (FX), thereby facilitating the activation of FX and promoting blood coagulation. Due to its prolonged half-life in patient plasma following administration, accurate detection of Emicizumab is clinically important. Enzyme-linked immunosorbent assay (ELISA) is a valuable method for monitoring Emicizumab levels, but it requires proteins that bind with high affinity to the antibody.

In this study, we engineered and expressed a recombinant fragment of Factor X containing the EGF-like 2 domain, designed to bind Emicizumab. A gene fragment encoding the EGF-like 2 domain of Factor X, previously identified as an Emicizumab-binding region, was codon-optimized and cloned into pUC57, then subcloned into the pET28a expression vector. Escherichia coli DH5α and E. coli Shuffle T7 (DE3) strains were used as cloning and expression hosts, respectively. The recombinant protein was purified and verified using 10 % SDS-PAGE, Bradford assay, and Western blot. Following expression, the recombinant protein was purified and quantified, with concentrations of 119 μg/mL for native and 81 μg/mL for denatured protein. These values correspond to total recoveries of 255 μg and 65 μg, respectively, from 50 mL cultures. These findings validate the use of this bacterially expressed FX-derived protein as a candidate component for incorporation into ELISA-based platforms aimed at Emicizumab detection.

Emicizumab是一种用于治疗a型血友病的治疗性抗体，它通过桥接活化因子IX （FXa）和因子X （FX）来模拟凝血因子VIII的功能，从而促进FX的活化并促进血液凝固。由于给药后其在患者血浆中的半衰期延长，因此准确检测Emicizumab在临床上非常重要。酶联免疫吸附试验（ELISA）是监测Emicizumab水平的一种有价值的方法，但它需要与抗体高亲和力结合的蛋白质。在这项研究中，我们设计并表达了含有egf样2结构域的重组因子X片段，旨在结合Emicizumab。编码因子X的egf样2结构域的基因片段先前被确定为emicizumab结合区，通过密码子优化并克隆到pUC57中，然后亚克隆到pET28a表达载体中。以大肠杆菌DH5α和大肠杆菌Shuffle T7 （DE3）菌株分别作为克隆和表达宿主。重组蛋白通过10% SDS-PAGE、Bradford assay和Western blot进行纯化和验证。表达后，对重组蛋白进行纯化和定量，原蛋白浓度为119 μg/mL，变性蛋白浓度为81 μg/mL。这些值对应于50ml培养物的总回收率分别为255 μg和65 μg。这些发现证实了这种细菌表达的fx衍生蛋白作为一种候选成分，可以结合到基于elisa的平台中，用于Emicizumab检测。

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引用次数: 0

Expression, purification and functional validation of a cancer-associated isoform of the HBx protein from human hepatitis B virus 人乙型肝炎病毒HBx蛋白癌症相关亚型的表达、纯化和功能验证

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification

Pub Date : 2026-02-01 Epub Date: 2025-11-03 DOI: 10.1016/j.pep.2025.106842

Alexis Clavier , Santiago Gómez-Evain , Toshinobu Shida , Rubaba R. Abanti , Franziska Hammerstein , Pavel Kielkowski , Mila Leuthold , Anne K. Schütz

The human hepatitis B virus (HBV) causes hepatitis B, a liver infection that can be acute or chronic. HBV encodes four proteins, among which the X protein (HBx) plays a critical role in viral replication. During chronic HBV infection, in which the viral DNA is integrated into the host genome, the HBx_1-120 isoform, comprising the N-terminal 120 residues, is highly expressed. Here, we describe a protocol for the recombinant overexpression and purification of untagged HBx_1-120 from bacterial cells. The procedure is compatible with stable isotope labelling in minimal media. Following cell lysis, HBx_1-120 was recovered from inclusion bodies (IBs), solubilized in urea, and purified by ion-exchange (IEX) and size-exclusion chromatography (SEC). The purified protein was extensively characterized, including by mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. Functionality was confirmed by a pulldown assay with a known interacting partner, Spindlin1. This protocol provides a robust framework to obtain untagged HBx_1-120 for structural and functional in vitro studies.

人类乙型肝炎病毒（HBV）引起乙型肝炎，这是一种急性或慢性肝脏感染。HBV编码四种蛋白，其中X蛋白（HBx）在病毒复制中起关键作用。在慢性HBV感染期间，病毒DNA被整合到宿主基因组中，包含n端120残基的HBx1-120亚型被高度表达。在这里，我们描述了一种从细菌细胞中重组过表达和纯化未标记HBx1-120的方案。该方法与稳定同位素标记在最小介质中兼容。细胞裂解后，从包涵体（IBs）中回收HBx1-120，在尿素中溶解，并通过离子交换（IEX）和尺寸排除色谱（SEC）纯化。纯化后的蛋白被广泛地表征，包括质谱和核磁共振（NMR）光谱。功能通过与已知的相互作用伙伴Spindlin1的下拉试验确认。该方案为获得用于体外结构和功能研究的未标记HBx1-120提供了一个强大的框架。

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引用次数: 0

Protein production in transgenic chickens mediated through primordial germ cells and embryonic stem cells 通过原始生殖细胞和胚胎干细胞介导转基因鸡的蛋白质生产。

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification

Pub Date : 2026-02-01 Epub Date: 2025-10-20 DOI: 10.1016/j.pep.2025.106828

Mengran Fang , Xinyue Chen , Yuan Liu, Hui Yuan

The inherent advantages of transgenic chickens, combined with the increasing demand for therapeutic proteins, are driving significant progress in transgenic chicken bioreactors. Primordial germ cells (PGCs) and embryonic stem cells (ESCs) are two pivotal carrier cells in the production of transgenic chicken bioreactors. Transgenic chickens have successfully produced therapeutic proteins using genetically modified and transferred PGCs. Meanwhile, considerable progress has been made in expressing therapeutic proteins using exogenous gene integration mediated by ESCs. Both carrier cells facilitate the efficient process of transgenic chickens. Here, we reviewed the characteristics, current applications, comparisons, and future directions of these two carrier cells, providing theoretical foundations and practical guidance for advancing development in the fields of chicken bioreactors.

转基因鸡的固有优势，加上对治疗性蛋白需求的不断增长，推动了转基因鸡生物反应器的重大进展。原始生殖细胞（PGCs）和胚胎干细胞（ESCs）是转基因鸡生物反应器生产中的两种关键载体细胞。转基因鸡已经成功地利用转基因和转移的PGCs生产出治疗性蛋白质。与此同时，利用ESCs介导的外源基因整合表达治疗性蛋白也取得了相当大的进展。这两种载体细胞促进了转基因鸡的高效过程。本文综述了这两种载体细胞的特点、应用现状、比较及未来发展方向，为推进鸡生物反应器领域的发展提供理论基础和实践指导。

引用次数: 0

Effect of culture media and fermentation process on the refolding and purification of rh-GCSF 培养基和发酵工艺对rh-GCSF再折叠和纯化的影响。

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification

Pub Date : 2026-02-01 Epub Date: 2025-10-21 DOI: 10.1016/j.pep.2025.106837

Somayeh Abolghasemi , Fatemeh Poureini , Valiollah Babaeipour , Faezeh Farji , Mohammad Reza Mofid

Human granulocyte-colony stimulating factor (h-GCSF) is used to mitigate neutropenia caused by chemotherapy. E. coli is the most common host for h-GCSF production due to its high yield and versatile culture strategies. However, overproduction of h-GCSF in E. coli often results in the formation of inclusion bodies (IBs) in the cytoplasm. The efficiency of refolding and purifying recombinant h-GCSF (rh-GCSF) produced as IBs depends on the expression strategy, induction, and cell growth conditions. This study investigated how different culture media and fermentation processes affect the refolding of IBs and the purification of rh-GCSF. The purified samples were compared to Neupogen® and PDgrastim reference standards using Western blotting, size exclusion chromatography, and reverse-phase high-performance liquid chromatography. The analysis confirmed that all proteins were correctly refolded and exhibited a purity exceeding 90 % after purification. The highest protein recovery percentage and purity of rh-GCSF with the lowest endotoxin content was achieved using M9 media in fed-batch culture.

人粒细胞集落刺激因子（h-GCSF）用于减轻化疗引起的中性粒细胞减少症。大肠杆菌是生产h-GCSF最常见的宿主，因为它的高产和多样化的培养策略。然而，大肠杆菌中h-GCSF的过量产生经常导致细胞质中包涵体（IBs）的形成。重组h-GCSF （rh-GCSF）以IBs的形式产生，其重折叠和纯化的效率取决于表达策略、诱导和细胞生长条件。本研究考察了不同培养基和发酵工艺对IBs再折叠和rh-GCSF纯化的影响。纯化后的样品与Neupogen®和PDgrastim标准品进行Western blotting、排色层析和反相高效液相色谱比较。分析证实所有蛋白都正确折叠，纯化后纯度超过90%。采用M9培养基进行分批补料培养，获得的rh-GCSF蛋白回收率和纯度最高，内毒素含量最低。

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引用次数: 0

Optimized Laccase production from the white rot fungi Pleurotus ostreatus and Trametes versicolor 白腐菌平菇和彩板菌产漆酶的优化。

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification

Pub Date : 2026-01-01 Epub Date: 2025-09-11 DOI: 10.1016/j.pep.2025.106813

Apoorva Deshmukh , Parnal Sattikar , Aishwarya Sukhatankar , Geetanjali Wakade , Pramod Kumbhar , Phaneeswara-Rao Kommoju

Laccases are predominantly found in bacteria, fungi, plants, and insects, and they have numerous industrial and biotechnological applications. Laccases are utilized in the pharmaceutical, food, pulp, and paper industries, and their cost-effective production in submerged culture conditions is of significant commercial value. Attempts were made to overexpress three laccase isoforms from Trametes versicolor (TV) in heterologous systems. Recombinant TV laccases were either insoluble in E.coli or poorly expressed in Pichia pastoris. Hence a submerged fermentation process was developed to produce these commercially important laccases from two non-recombinant white rot fungi: Pleurotus ostreatus (PO) and Trametes versicolor (TV). Molasses and corn steep liquor (CSL) were used as carbon and nitrogen sources, respectively, while 2,5-xylidine, sodium lignosulfonate, and copper sulfate were used as inducers, making the entire process economical. Laccase activity reached a maximum of 374,000 U/L (or 374 kU/L) in 20 days. When evaluated at 45 °C, TV laccase outperformed PO laccase in terms of stability, a crucial factor in the delignification of biomass. TV laccase demonstrated superior stability over PO laccase at 45 °C, making it the preferred choice for biomass pre-treatment applications. We demonstrate the use of this laccase in two industrial applications.

1. Lignocellulose Depolymerization: Treatment of rice straw with Pleurotus ostreatus laccase resulted in visible structural distortion, as observed under a scanning electron microscope.

2. Industrial Wastewater Treatment: Significant decolorization of indigo carmine and Remazol Brilliant Blue dyes was achieved overnight, with reductions of up to 70 % and 74 %, respectively, when incubated with Trametes versicolor laccase.

漆酶主要存在于细菌、真菌、植物和昆虫中，它们在工业和生物技术上有许多应用。漆酶被用于制药、食品、纸浆和造纸工业，它们在水下培养条件下的成本效益生产具有重要的商业价值。尝试在异源系统中过表达三种色板菌漆酶同工型。重组TV漆酶要么在大肠杆菌中不溶，要么在毕赤酵母中表达不良。因此，研究人员利用两种非重组白腐菌Pleurotus ostreatus （PO）和Trametes versicolor （TV）开发了一种深层发酵工艺，以生产这些具有重要商业价值的漆酶。以糖蜜和玉米浆（CSL）分别为碳源和氮源，以2,5-二甲醚、木质素磺酸钠和硫酸铜为诱导剂，使整个过程具有经济性。漆酶活性在20 d内达到最大值374,000 U/L（或374 kU/L）。当在45°C下评估时，TV漆酶在稳定性方面优于PO漆酶，稳定性是生物质脱木质素的关键因素。TV漆酶在45°C下表现出比PO漆酶更好的稳定性，使其成为生物质预处理应用的首选。我们演示了这种漆在两个工业应用中的使用：1。木质纤维素解聚：在扫描电子显微镜下观察到，用平菇漆酶处理稻草导致明显的结构扭曲。2. 工业废水处理：靛蓝胭脂红和Remazol Brilliant Blue染料在一夜之间实现了显著的脱色，当与Trametes versicolor漆酶孵育时，脱色率分别高达70%和74%。

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引用次数: 0

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 Q4 BIOCHEMICAL RESEARCH METHODS

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

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 (MnCl₂) 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.

为了确保生物类似药和参比药之间的高度相似性，进行全面的表征和严格控制糖基化过程至关重要。在这里，我们旨在优化双特异性抗体（BsAb）的生物类似物的糖基化谱，通过糖基化调节剂的协同作用，使其与参比药物非常相似。为了确定最强的调节剂和合适的浓度范围，我们首先研究了不同浓度的半乳糖（Gal）和氯化锰（MnCl2）对摇瓶中半乳糖基化速率的影响，以及三（羟甲基）氨基甲烷（tris）对甘露糖掺入和2 L生物反应器中的影响。重要的是，Tris和半乳糖同时使用不会对n -聚糖修饰产生任何相互作用，也不会对细胞生长、代谢、抗体电荷变异或纯度产生有害影响。综上所述，同时使用0.75 mM Tris和8 mM半乳糖可以产生Bs-mAb1的糖基化谱，这与参考药物的糖基化谱高度相似，从而为优化生物类似药的糖基化提供了有效的策略。这些发现为治疗性单克隆抗体生产中糖基化的调控提供了重要的见解，并可能有助于提高生物仿制药的一致性和治疗性能。

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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 Q4 BIOCHEMICAL RESEARCH METHODS

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

The plant disease myrtle rust is caused by the fungus Austropuccinia psidii. It has led to functional myrtaceous species extinctions in Australia and is a significant threat to other species globally. During infection, A. psidii 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 A. psidii effector proteins were recombinantly produced using SHuffle Escherichia coli 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 A. psidii protein provides a foundation for future investigation of the molecular mechanisms of A. psidii pathogenicity.

桃金娘锈病是一种由桃金娘锈病真菌引起的植物病害。它已导致澳大利亚功能性桃金娘科物种灭绝，并对全球其他物种构成重大威胁。在感染期间，psidii分泌效应蛋白来操纵宿主植物的防御。在这种病原体的基因组中编码了许多可能的效应物，其中一些在芽孢萌发和植物组织初始入侵的早期表达。4种推测的效应蛋白（AP1260、AP5292、AP10948和AP143）在感染后24-48小时内差异表达，提示它们在感染过程中发挥重要作用。与其他锈菌一样，这些效应蛋白很小且富含半胱氨酸，通常形成二硫键，分离它们进行生物物理表征可能具有挑战性。AlphaFold3模型预测AP1260、AP5292、AP10948和AP143形成二硫键，而无序分析表明存在内在无序区域。利用重组大肠杆菌细胞和适应性共表达载体“ApFunCyDisCo”重组产生了四种推测的psidii效应蛋白。成功制备了三种效应器，但不溶性。第四种效应器AP1260成功地从可溶性部分中产生，并通过四步工艺纯化：固定化金属亲和层析、脱盐、阴离子交换层析和尺寸排除层析。圆二色光谱分析表明，AP1260主要具有无规螺旋结构，但同时含有β-链和α-螺旋结构。这一首次成功的psidii蛋白的制备和分离为psidii致病性分子机制的进一步研究奠定了基础。

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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 Q4 BIOCHEMICAL RESEARCH METHODS

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

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 Escherichia coli. 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.

C14orf119是一种功能未知的线粒体蛋白，可能与缺血性卒中发病机制有关。为了进行全面的生物学研究，我们开发并验证了针对该靶点的特异性多克隆抗体。我们的方法包括在大肠杆菌中对His-和gst标记的C14orf119进行原核表达和层析纯化。纯化后的His-C14orf119作为兔多克隆抗体生产的免疫原，纯化后的GST-C14orf119用于随后的目标特异性抗体亲和纯化。严格的鉴定表明该抗体对外源性和内源性C14orf119都具有特异性。值得注意的是，C14orf119抗体未能检测到对照his标记的蛋白，证实了在基于gst的亲和纯化过程中有效去除了抗his抗体。这种经过严格验证的抗体为C14orf119的功能表征提供了关键的分子工具，为其病理生理作用的机制研究铺平了道路。

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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 Q4 BIOCHEMICAL RESEARCH METHODS

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

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 Saccharomyces cerevisiae 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 (OD₆₀₀ ≈ 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.

猪表皮生长因子（PoEGF）促进断奶仔猪肠上皮发育，增强免疫功能，有可能作为饲料中抗生素的替代品。然而，高生产成本和复杂的纯化要求阻碍了重组PoEGF （rPoEGF）的大规模应用。本研究通过整合α-因子信号peptide-PoEGF-6×His表达盒，构建了一株能够表达和分泌rPoEGF的重组酿酒酵母INVSc1菌株。通过SDS-PAGE和Western blot分析证实了表达，并通过BALB/c 3T3成纤维细胞增殖试验验证了生物活性，产生的最大有丝分裂活性为10,963 U/mL。为了提高生产效率，在4-L生物反应器中开发并优化了两阶段发酵策略。该过程包括葡萄糖为基础的分批补料生长阶段，然后在优化条件（25°C, pH 5.0, DO 30%）下进行半乳糖诱导表达。该方法实现了高密度培养（OD600≈71.5），最终分泌蛋白产量为30.2 mg/L。值得注意的是，含有活性rPoEGF的培养物可以直接用作功能性饲料添加剂，而无需进行下游纯化。这些结果为生产具有生物活性的rPoEGF提供了一种实用的、可扩展的、具有成本效益的方法，并支持其作为一种安全的、无抗生素的生长促进剂在猪生产中的应用。

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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 Q4 BIOCHEMICAL RESEARCH METHODS

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

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.

继首次报道小鼠诱导多能干细胞（iPS）后，已经建立了各种哺乳动物来源的iPS细胞。相比之下，来自禽类的iPS细胞或iPS样细胞鲜有报道。iPS细胞可以分化成各种细胞类型（如神经细胞和肝细胞），并在培养中无限增殖。与胚胎干细胞不同，诱导多能干细胞是由体细胞产生的，因此不需要胚胎。由于体细胞可以从死亡个体中获得，iPS细胞技术可以适用于野生鸟类物种，而不仅仅是在鸡身上的应用。本小组之前报道了利用修饰的八聚体结合转录因子3/4 （Oct3/4）、sly -box转录因子2 （Sox2）、kr ppel样因子4 （Klf4）、MYC原癌基因（c-Myc）、Nanog和lin-28同源物a （Lin28A）从体细胞中生成鸡iPS细胞。发育中的鸡蛋是生产蛋白质的宝贵资源。我们可以使用基因组编辑或转基因鸡iPS细胞从鸡的嵌合发育蛋中获得有价值的蛋白质。此外，我们的团队利用修饰过的- oct3 /4、Sox2、Klf4、c-Myc、Nanog、Lin28和Klf2建立了从濒危鸟类（冲绳秧鸡、日本石鸡和Blakiston鱼鸮）中提取的iPS细胞。从诱导多能干细胞分化出来的细胞（如神经细胞和肝细胞）可用于兽医学的药物测试和评估对传染病和污染物的敏感性。我们相信iPS细胞技术可以发展成为保护濒危鸟类物种的有力工具。

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引用次数: 0