Protein expression and purification最新文献_第10页

Unravelling the reason for different binding behaviors exhibited by antibody aggregates towards preparative and analytical Protein A columns 揭示抗体聚集体与制备型和分析型蛋白 A 柱表现出不同结合行为的原因。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-27 DOI: 10.1016/j.pep.2024.106449

Wanyuan Dong, Mengying Tian, Yifeng Li

{"title":"Unravelling the reason for different binding behaviors exhibited by antibody aggregates towards preparative and analytical Protein A columns","authors":"Wanyuan Dong, Mengying Tian, Yifeng Li","doi":"10.1016/j.pep.2024.106449","DOIUrl":"10.1016/j.pep.2024.106449","url":null,"abstract":"<div><p>We previously showed that the root cause of low Protein A step yield observed for certain antibodies/Fc-fusions is the presence of non-binding aggregates in cell culture harvest. A pre-assumption for the above conclusion is that the aggregates, while do not bind to the preparative Protein A column, can bind to the analytical Protein A-high performance liquid chromatography (HPLC) column used for titer measurement. In the current work, using materials from a previous case with the low yield issue, we confirmed that non-binding aggregates in preparative Protein A flow-through can indeed bind to the analytical Protein A column. In addition, we showed that this discrepancy is mainly due to the different loading densities applied under these two circumstances. We also demonstrated that aggregate bound to the analytical Protein A column slightly stronger than the monomer, as it exhibited a longer retention time. In summary, the current study not only confirmed that non-binding aggregates detected in the preparative Protein A flow-through bind to the Protein A-HPLC column and contribute to the measured titer of culture harvest but also unravelled the reason for different binding behaviors exhibited by antibody aggregates towards preparative and analytical Protein A columns.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139997235","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

Adapting recombinant bacterial alkaline phosphatase for nucleotide exchange of small GTPases 将重组细菌碱性磷酸酶用于小 GTP 酶的核苷酸交换。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-22 DOI: 10.1016/j.pep.2024.106446

Peter H. Frank, Min Hong, Brianna Higgins, Shelley Perkins, Troy Taylor, Vanessa E. Wall, Matthew Drew, Timothy Waybright, William Gillette, Dominic Esposito, Simon Messing

引用次数: 0

Effective expression and characterization of the receptor binding domains in SARS-CoV-2 Spike proteins from original strain and variants of concern using Bombyx mori nucleopolyhedrovirus in silkworm 利用蚕核型多角体病毒有效表达和鉴定 SARS-CoV-2 Spike 蛋白原始株和关注变异株的受体结合域。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-22 DOI: 10.1016/j.pep.2024.106450

Akira Tsukamoto , Lee Jae Man , Kosuke Oyama , Akitsu Masuda , Hiroaki Mon , Tadashi Ueda , Takahiro Kusakabe

{"title":"Effective expression and characterization of the receptor binding domains in SARS-CoV-2 Spike proteins from original strain and variants of concern using Bombyx mori nucleopolyhedrovirus in silkworm","authors":"Akira Tsukamoto , Lee Jae Man , Kosuke Oyama , Akitsu Masuda , Hiroaki Mon , Tadashi Ueda , Takahiro Kusakabe","doi":"10.1016/j.pep.2024.106450","DOIUrl":"10.1016/j.pep.2024.106450","url":null,"abstract":"<div><p>A new coronavirus, known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the global pandemic of COVID-19 in 2020. Through structural analysis, it was found that several amino acid residues in the human angiotensin-converting enzyme-2 (hACE2) receptor directly interact with those in the receptor binding domain (RBD) of the spike glycoprotein (S-protein). Various cell lines, including HEK293, HeLa cells, and the baculovirus expression vector system (BEVS) with the insect cell line Sf9, have been utilized to produce the RBD. In this study, we investigated the use of <em>Bombyx mori</em> nucleopolyhedrovirus (BmNPV) and BEVS. For efficient production of a highly pure recombinant RBD protein, we designed it with two tags (His tag and STREP tag) at the C-terminus and a solubilizing tag (SUMO) at the N-terminus. After expressing the protein using BmNPV and silkworm and purifying it with a HisTrap excel column, the eluted protein was digested with SUMO protease and further purified using a Strep-Tactin Superflow column. As a result, we obtained the RBD as a monomer with a yield of 2.6 mg/10 mL serum (equivalent to 30 silkworms). The RBD showed an affinity for the hACE2 receptor. Additionally, the RBDs from the Alpha, Beta, Gamma, Delta, and Omicron variants were expressed and purified using the same protocol. It was found that the RBD from the Alpha, Beta, Gamma, and Delta variants could be obtained with yields of 1.4–2.6 mg/10 mL serum and had an affinity to the hACE2 receptor.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139940699","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

Surface display system of Bacillus subtilis: A promising approach for improving the stability and applications of cellobiose dehydrogenase 枯草芽孢杆菌表面展示系统：提高纤维生物糖脱氢酶的稳定性和应用前景广阔的方法。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-17 DOI: 10.1016/j.pep.2024.106448

Zhengfen Wu, Pengfei Li, Xihua Chen, Yong Feng, Yi Ma, Zhong Ni, Daochen Zhu, Huayou Chen

{"title":"Surface display system of Bacillus subtilis: A promising approach for improving the stability and applications of cellobiose dehydrogenase","authors":"Zhengfen Wu, Pengfei Li, Xihua Chen, Yong Feng, Yi Ma, Zhong Ni, Daochen Zhu, Huayou Chen","doi":"10.1016/j.pep.2024.106448","DOIUrl":"10.1016/j.pep.2024.106448","url":null,"abstract":"<div><p>Cellobiose dehydrogenase (CDH) plays a crucial role in lignocellulose degradation and bioelectrochemical industries, making it highly in demand. However, the production and purification of CDH through fungal heterologous expression methods is time-consuming, costly, and challenging. In this study, we successfully displayed <em>Pycnoporus sanguineus</em> CDH (<em>ps</em>CDH) on the surface of <em>Bacillus subtilis</em> spores for the first time. Enzymatic characterization revealed that spore surface display enhanced the tolerance of <em>ps</em>CDH to high temperature (80 °C) and low pH levels (3.5) compared to free <em>ps</em>CDH. Furthermore, we found that glycerol, lactic acid, and malic acid promoted the activity of immobilized spore-displayed <em>ps</em>CDH; glycerol has a more significant stimulating effect, increasing the activity from 16.86 ± 1.27 U/mL to 46.26 ± 3.25 U/mL. After four reuse cycles, the <em>ps</em>CDH immobilized with spores retained 48% of its initial activity, demonstrating a substantial recovery rate. In conclusion, the spore display system, relying on cotG, enables the expression and immobilization of CDH while enhancing its resistance to adverse conditions. This system demonstrates efficient enzyme recovery and reuse. This approach provides a novel method and strategy for the immobilization and stability enhancement of CDH.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139906303","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 performance production process development and scale-up of an anti-TSLP nanobody 高效生产工艺的开发和抗 TSLP 纳米抗体的放大。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-16 DOI: 10.1016/j.pep.2024.106441

Xiaofei Li, Peng Qiao, Yicai Zhang, Guoxin Liu, Min Zhu, Junwei Gai, Yakun Wan

{"title":"High performance production process development and scale-up of an anti-TSLP nanobody","authors":"Xiaofei Li, Peng Qiao, Yicai Zhang, Guoxin Liu, Min Zhu, Junwei Gai, Yakun Wan","doi":"10.1016/j.pep.2024.106441","DOIUrl":"10.1016/j.pep.2024.106441","url":null,"abstract":"<div><p>Nanobodies (Nbs) represent a class of single-domain antibodies with great potential application value across diverse biotechnology fields, including therapy and diagnostics. Thymic Stromal Lymphopoietin (TSLP) is an epithelial cell-derived cytokine, playing a crucial role in the regulation of type 2 immune responses at barrier surfaces such as skin and the respiratory/gastrointestinal tract. In this study, a method for the expression and purification of anti-TSLP nanobody (Nb3341) was established at 7 L scale and subsequently scaled up to 100 L scale. Key parameters, including induction temperature, methanol feed and induction pH were identified as key factors by Plackett-Burman design (PBD) and were optimized in 7 L bioreactor, yielding optimal values of 24 °C, 8.5 mL/L/h and 6.5, respectively. Furthermore, Diamond Mix-A and Diamond MMC were demonstrated to be the optimal capture and polishing resins. The expression and purification process of Nb3341 at 100L scale resulted in 22.97 g/L titer, 98.7% SEC-HPLC purity, 95.7% AEX-HPLC purity, 4 ppm of HCP content and 1 pg/mg of HCD residue. The parameters of the scaling-up process were consistent with the results of the optimized process, further demonstrating the feasibility and stability of this method. This study provides a highly promising and competitive approach for transitioning from laboratory-scale to commercial production-scale of nanobodies.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139898209","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

Backbone cyclization of Salmonella typhimurium diaminopropionate ammonia-lyase to enhance the activity and stability 鼠伤寒沙门氏菌二氨基丙酸氨水解酶的骨架环化以提高其活性和稳定性。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-16 DOI: 10.1016/j.pep.2024.106447

Xiaomei He , Tingting Lin , Yuying Xie , Jinjing Li , Yuanyuan Ge , Shuncheng Zhang , Jun Fan

{"title":"Backbone cyclization of Salmonella typhimurium diaminopropionate ammonia-lyase to enhance the activity and stability","authors":"Xiaomei He , Tingting Lin , Yuying Xie , Jinjing Li , Yuanyuan Ge , Shuncheng Zhang , Jun Fan","doi":"10.1016/j.pep.2024.106447","DOIUrl":"10.1016/j.pep.2024.106447","url":null,"abstract":"<div><p>Diaminopropionate ammonia-lyase transforms D and L isomers of 2,3-diaminopropionate to pyruvate and ammonia. It catalyzes D- and <span>l</span>-serine less effectively. L-2,3-diaminopropionate is a precursor in the biosynthesis of oxalyl diaminopropionate as a neurotoxin in certain legume species. In this work, we cyclized the diaminopropionate ammonia-lyase from <em>Salmonella typhimurium</em> in vitro using the redox-responsive split intein, and identified that backbone cyclization afforded the enzyme with the improved activity, thermal stability and resistance to the exopeptidase proteolysis, different from effects of the incorporated sequence recognized by tobacco vein mottling virus protease at <em>C-</em>terminus. Using analyses of three fluorescent dyes including 8-anilino-1-naphthalenesulfonic acid, N-phenyl-1-naphthylamine, and thioflavin T, the same amounts of the cyclic protein displayed less fluorescence than those of the linear protein upon the heat treatment. The cyclic enzyme displayed the enhanced activity in <em>Escherichia coli</em> cells using the designed novel reporter. In this system, <span>d</span>-serine was added to the culture and transported into the cytoplasm. It was transformed by pre-overexpression of the diaminopropionate ammonia-lyase, and untransformed <span>d</span>-serine was oxidized by the coproduced human <span>d</span>-amino acid oxidase to generate hydrogen peroxide. This oxidant is monitored by the HyPer indicator. The current results presented that the cyclized enzyme could be applied as a better candidate to block the neurotoxin biosynthesis in certain plant species.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139900330","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

The expression of HIV-1 tat in Lactococcus lactis 乳酸乳球菌中 HIV-1 tat 的表达。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-14 DOI: 10.1016/j.pep.2024.106443

Deepak Selvam , Anish D'silva , Arun Panchapakesan , Yuvrajsinh Gohil , Jayendra Singh , Luke Elizabeth Hanna , Udaykumar Ranga

引用次数: 0

Novel melanin-derived stationary phase for immobilized metal ion affinity chromatography in recombinant His-tagged protein purification 新型黑色素固定相，用于重组 His 标记蛋白质纯化中的固定金属离子亲和层析。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-14 DOI: 10.1016/j.pep.2024.106444

Hong-Nhung Le Thi , Ngoc-Tram Le , Thu-Hoai Bui Thi , Hong-Loan Nguyen Thi , Thanh-Thuy Nguyen , Yen Nguyen Thi , Minh-Ngoc Ha , Dinh-Thang Nguyen

{"title":"Novel melanin-derived stationary phase for immobilized metal ion affinity chromatography in recombinant His-tagged protein purification","authors":"Hong-Nhung Le Thi , Ngoc-Tram Le , Thu-Hoai Bui Thi , Hong-Loan Nguyen Thi , Thanh-Thuy Nguyen , Yen Nguyen Thi , Minh-Ngoc Ha , Dinh-Thang Nguyen","doi":"10.1016/j.pep.2024.106444","DOIUrl":"10.1016/j.pep.2024.106444","url":null,"abstract":"<div><p>The matrix of the stationary phase is a crucial element in affinity chromatography for protein purification. Various materials, including polymer or magnetic materials, have been employed as the matrix in the purification of His-tagged protein. Here, for the first time, we utilized a combination of melanin and alginate, both natural polymer materials, to synthesize Ni-melanin/alginate (Ni-M/A) beads for His-tagged protein purification. We investigated the binding of His-tagged Mpro on the Ni-M/A beads, referred to as Ni-M/A-Mpro, and assessed the elution efficiency of Mpro from the beads. Our examination involved FTIR, EDS, XRD, SDS-PAGE, and Western blotting methods. FTIR spectra revealed notable changes in the stretching patterns and intensities of hydroxyl, amine, carbonyl, imine and amide chemical groups, when Mpro protein was present in the Ni-M/A sample. XRD spectra demonstrated the occurrence of two Nickel peaks at 35–40 deg and 40–45 deg in Ni-M/A, but only one nickel peak at 35–40 deg in Ni-M/A-Mpro, indicating the binding of Mpro on the Nickel ions. EDS analysis reported a decrease in the concentration of Nickel on the surface of Ni-M/A from 16% to 7% when Mpro protein was loaded into the stationary phase. Importantly, our data indicated that the purity of the His-tagged protein Mpro after purification reached 97% after just one-step purification using the Ni-M/A stationary phase. Moreover, the binding capacity of Ni-M/A for Mpro was approximately 5.2 mg/g with recovery efficiency of 40%. Our results suggested Ni-M/A as a highly potential solid phase for affinity chromatography in the purification of His-tagged protein.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139747297","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

Evaluation of the paired-Cas9 nickase and RNA-guided FokI genome editing tools in precise integration of an anti-CD52 bicistronic monoclonal antibody expression construct at Chinese hamster ovary cells 18S rDNA locus 在中国仓鼠卵巢细胞 18S rDNA 基因座上精确整合抗 CD52 双单克隆抗体表达构建体时，对成对 Cas9 快速酶和 RNA 引导的 FokI 基因组编辑工具的评估。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-09 DOI: 10.1016/j.pep.2024.106445

Hadi Bayat , Faranak Farahmand , Sayed Hassan Tabatabaee , Forough Shams , Omid Mohammadian , Es'hagh Pourmaleki , Azam Rahimpour

{"title":"Evaluation of the paired-Cas9 nickase and RNA-guided FokI genome editing tools in precise integration of an anti-CD52 bicistronic monoclonal antibody expression construct at Chinese hamster ovary cells 18S rDNA locus","authors":"Hadi Bayat , Faranak Farahmand , Sayed Hassan Tabatabaee , Forough Shams , Omid Mohammadian , Es'hagh Pourmaleki , Azam Rahimpour","doi":"10.1016/j.pep.2024.106445","DOIUrl":"10.1016/j.pep.2024.106445","url":null,"abstract":"<div><h3>Introduction</h3><p>The aim of this study was to compare two CRISPR/Cas9-based orthogonal strategies, paired-Cas9 nickase (paired-Cas9n) and RNA-guided FokI (RFN), in targeting 18S rDNA locus in Chinese hamster ovary (CHO) cells and precisely integrating a bicistronic anti-CD52 monoclonal antibody (mAb) expression cassette into this locus.</p></div><div><h3>Methods</h3><p>T7E1 and high-resolution melt (HRM) assays were used to compare the ability of mentioned systems in inducing double-strand break (DSB) at the target site. Moreover, 5′- and 3′-junction polymerase chain reactions (PCR) were used to verify the accuracy of the targeted integration of the mAb expression cassette into the 18S rDNA locus. Finally, anti-CD52 mAb gene copy number was measured and, its expression was analyzed using ELISA and western blot assays.</p></div><div><h3>Results</h3><p>Our results indicated that both paired-Cas9n and RFN induced DSB at the target site albeit RFN performance was slightly more efficient in HRM analysis. We also confirmed that the anti-CD52 mAb cassette was accurately integrated at the 18S rDNA locus and the mAb was expressed successfully in CHO cells.</p></div><div><h3>Conclusion</h3><p>Taken together, our findings elucidated that both paired-Cas9n and RFN genome editing tools are promising in targeting the 18S rDNA locus. Site specific integration of the bicistronic anti-CD52 mAb expression cassette at this locus in the CHO-K1 cells was obtained, using RFN. Moreover, proper expression of the anti-CD52 mAb at the 18S rDNA target site can be achieved using the bicistronic internal ribosome entry site (IRES)-based vector system.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139717774","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

Scalable dual column cation exchange affinity chromatography based platform process for recombinant protein purification 基于双柱阳离子交换亲和层析平台的可扩展重组蛋白纯化工艺。

IF 1.6 4区生物学

Protein expression and purification Pub Date : 2024-02-07 DOI: 10.1016/j.pep.2024.106442

Sai Vivek Prabhala, David W. Wood

{"title":"Scalable dual column cation exchange affinity chromatography based platform process for recombinant protein purification","authors":"Sai Vivek Prabhala, David W. Wood","doi":"10.1016/j.pep.2024.106442","DOIUrl":"10.1016/j.pep.2024.106442","url":null,"abstract":"<div><p>A novel tandem affinity tag is presented that enables the use of cation exchange resins for initial affinity purification, followed by an additional column step for enhanced purity and affinity tag self-removal. In this method, the highly charged heparin-binding tag binds strongly and selectively to either a strong or weak cation exchange resin based on electrostatic interactions, effectively acting as an initial affinity tag. Combining the heparin-binding tag (HB-tag) with the self-removing <em>i</em>CapTag™ provides a means for removing both tags in a subsequent self-cleaving step. The result is a convenient platform for the purification of diverse tagless proteins with a range of isoelectric points and molecular weights. In this work, we demonstrate a dual column process in which the tagged protein of interest is first captured from an <em>E. coli</em> cell lysate using a cation exchange column via a fused heparin-binding affinity tag. The partially purified protein is then diluted and loaded onto an <em>i</em>CapTag™ split-intein column, washed, and then incubated overnight to release the tagless target protein from the bound tag. Case studies are provided for enhanced green fluorescent protein (eGFP), beta galactosidase (βgal), maltose binding protein (MBP) and beta lactamase (βlac), where overall purity and host cell DNA clearance is provided. Overall, the proposed dual column process is shown to be a scalable platform technology capable of accessing both the high dynamic binding capacity of ion exchange resins and the high selectivity of affinity tags for the purification of recombinant proteins.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2024-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1046592824000147/pdfft?md5=87d5ecd37bc37609f0d47b7dd53471f7&pid=1-s2.0-S1046592824000147-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139712901","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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