Sezen Demirhan-Yazıcı, Kemal Karaca, Ayşe Nalbantsoy, Rengin Eltem
{"title":"An economically cost-effective production medium optimization of Bacillus subtilis Ö-4-68: A potential probiotic for aquaculture.","authors":"Sezen Demirhan-Yazıcı, Kemal Karaca, Ayşe Nalbantsoy, Rengin Eltem","doi":"10.1002/btpr.70070","DOIUrl":"https://doi.org/10.1002/btpr.70070","url":null,"abstract":"<p><p>Probiotic use has become more important in aquaculture for healthy and sustainable output. In particular, Bacillus spp. have emerged as effective probiotic agents, improving gut health, enhancing the immune system, promoting growth, and providing protection against pathogens in fish. Therefore, the application of Bacillus in aquaculture offers a strategic approach to increasing productivity while reducing the reliance on antibiotics. In this study, the antibacterial activities of Bacillus isolates, whose probiotic properties will be determined, against test bacteria that are fish pathogens such as Aeromonas hydrophila, Vibrio anguillarum, Lactococcus garvieae, and Yersinia ruckeri were determined by using cross-streak method and agar well diffusion methods. Then, antibiotic resistances of 75 isolates determined to have antibacterial activity were screened against 9 different antibiotics by the agar disc diffusion method. Gastric juice (pH 2.5) tolerance of 55 isolates determined to be sensitive to antibiotics was examined, and the tolerance of 13 isolates to gastric juice was determined. Optimum growth characteristics at acidic pH, surface hydrophobicity, bile tolerance, and protease, amylase, lipase, and cellulase activities, hemolytic activities, coagulase activities, bacterial adhesion abilities, and biofilm production properties of these isolates were determined. As a result, Bacillus subtilis Ö-4-68, with the best probiotic properties, was selected from the examined isolates, and production medium optimization was carried out with laboratory scale statistical experiment design (Response Surface Methodology, RSM) for high amount of biomass production. As a result of the trials, an economical cost-effective production medium content with high biomass production was determined.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70070"},"PeriodicalIF":2.5,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145129977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mohd Asim Khan, Griffin J Beyer, Naomi Goosby, Lilly Ortiz, Andre F Palmer
{"title":"Scalable production and biophysical characterization of an enzyme cocktail derived from human red blood cells.","authors":"Mohd Asim Khan, Griffin J Beyer, Naomi Goosby, Lilly Ortiz, Andre F Palmer","doi":"10.1002/btpr.70072","DOIUrl":"https://doi.org/10.1002/btpr.70072","url":null,"abstract":"<p><p>Red blood cells (RBCs) play a critical role in oxygen and carbon dioxide transport, which is facilitated by RBC-encapsulated hemoglobin (Hb) and carbonic anhydrase (CA). In addition, RBCs are constantly exposed to oxidative stress due to the intracellular reactive oxygen species (ROS) generated during Hb auto-oxidation. Antioxidant enzymes within RBCs, such as superoxide dismutase (SOD), catalase (CAT), and peroxiredoxin (Prx), counteract ROS generation to protect the RBC from oxidative stress. Therefore, this study presents a scaled-up method to extract an enzyme cocktail from lysed human RBCs, enriched with the major RBC enzymes with minimal Hb contamination. Using ethanol-chloroform precipitation and multiple biophysical analyses (SDS-PAGE, SEC-HPLC, MALDI-TOF, and LC-MS/MS), the RBC enzymes were successfully separated from Hb in the hemolysate. The purified enzyme cocktail exhibited minimal Hb contamination and retained a significant amount of CA, and antioxidative enzymes like SOD and CAT. Therefore, this scalable RBC enzyme purification method provides an efficient approach for isolating RBC enzymes with broad biomedical relevance.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70072"},"PeriodicalIF":2.5,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145085167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Exploring the design space for Triton X-100 substitutes in viral inactivation applications.","authors":"Yuqi Du, Shanshan Wu","doi":"10.1002/btpr.70069","DOIUrl":"10.1002/btpr.70069","url":null,"abstract":"<p><p>The urgent need to replace the European-prohibited Triton X-100 in biomanufacturing has been hindered by insufficient data on alternative detergents' minimum effective concentrations (MECs) and process robustness in viral inactivation. This study makes systematic research including: (1) Establishment of MECs for novel Triton X-100 substitutes (TXR-1/VIS/13-S9/C16) achieving effective inactivation of Xenotropic murine leukemia virus and Pseudorabies virus (log<sub>10</sub> reduction factor >4) across diverse CHO harvest fluids; (2) Demonstration of broad-spectrum efficacy against various viruses, with TXR-1/VIS/13-S9 maintaining effective inactivation for Bovine viral diarrhea virus, Vesicular stomatitis virus, Baculovirus, and Herpes simplex virus type 1; (3) Identification of PS20's material-dependent inactivation dynamics, establishing standalone parameters (4 h at 37°C) that achieve equivalent viral inactivation to traditional tri(n-butyl)phosphate -combined methods without requiring lipase activity-a paradigm shift in detergent application. Crucially, process optimization revealed that extending exposure time (1-4 h) enhanced PS20/PS80 efficacy more effectively than two fold concentration increases, providing cost-effective solutions. These findings deliver broader design spaces for implementing eco-friendly detergents while ensuring compliance with EMA/ICH viral safety standards.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70069"},"PeriodicalIF":2.5,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Roli Kargupta, Shannon Rivera, Brent Kochert, Kyle Devenney, Daniel Donelly, Tariq Atieh, Fang Li, Jessica Pan, Daya Patel, Venkata Tayi, Gaurav Chauhan, Rebecca Chmielowski, Susan J Abbondanzo
{"title":"Elucidation of cell culture impacts on hydroxylysine levels in monoclonal antibodies using high-throughput analytical quantification and media components.","authors":"Roli Kargupta, Shannon Rivera, Brent Kochert, Kyle Devenney, Daniel Donelly, Tariq Atieh, Fang Li, Jessica Pan, Daya Patel, Venkata Tayi, Gaurav Chauhan, Rebecca Chmielowski, Susan J Abbondanzo","doi":"10.1002/btpr.70068","DOIUrl":"https://doi.org/10.1002/btpr.70068","url":null,"abstract":"<p><p>Hydroxylysine (Hyl) is a post-translational hydroxyl modification of lysine that is not commonly observed at very high levels and thus is not usually considered a product quality attribute (PQA). Post-translation modifications (PTMs) are considered potential PQAs when elevated levels are observed - requiring monitoring and investigation. In a recent monoclonal antibody expression using Media A, Hyl levels were observed at ~20%-35%. At such elevated percentage levels, Hyl was considered a PQA - triggering a root-cause investigation in the upstream activities like cell culture conditions and media components. Initial detection of the Hyl modification originated from non-quantitative, intact mass analysis with confirmation of site-location determined by peptide mapping. Through the root-cause investigation, it was determined that levels of Hyl were underestimated by ~10-fold using tryptic peptide mapping analysis without inclusion of miscleaved peptides. The analytical procedure was revised from trypsin-digestion to IdeS-digestion, a reduced mass analysis, to accurately and rapidly quantify Hyl levels of investigational samples. Proprietary Media B was utilized to reduce the Hyl level by 2-fold to ~10%-15%. Further investigation into the media and feed components determined that increasing concentration of Fe(III) content decreased Hyl levels. Supplementation of Fe(III) served as a robust mitigation strategy of Hyl reduction in upstream process. Media B was used to scale up to a 500 L bioreactor while maintaining the lower Hyl level. The analytical and cell culture methods developed in this study can be leveraged to detect and tune Hyl levels.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70068"},"PeriodicalIF":2.5,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940777","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alice Delhaes, Laure Bataille, Myriam Médéric, Sébastien Lecommandoux, Elisabeth Garanger
{"title":"Production of recombinant methionine-containing elastin-like polypeptides in a fermenter using ECPM1 medium.","authors":"Alice Delhaes, Laure Bataille, Myriam Médéric, Sébastien Lecommandoux, Elisabeth Garanger","doi":"10.1002/btpr.70057","DOIUrl":"https://doi.org/10.1002/btpr.70057","url":null,"abstract":"<p><p>Elastin-like polypeptides (ELPs) are recombinant protein-like polymers whose macromolecular structure can be precisely controlled through genetic manipulation of their sequence and length. Their lower critical solution temperature (LCST) phase behavior facilitates purification via chromatography-free techniques and can be explored for self-assembly. As a result, ELPs are extensively investigated for diverse biological, biomedical, and biotechnological applications. So far, ELPs have mostly been isolated from bacteria grown in flasks or fermenters containing complex media that only yield limited amounts of biomass. We herein explored the use of the semi-defined ECPM1 medium, known to limit the accumulation of toxic metabolites and rich in glycerol as a low energy carbon source, to produce ELPs of different chain lengths and containing oxidation-sensitive methionine residues. We report the optimized bioproduction using ECPM1 of ELP[M<sub>1</sub>V<sub>3</sub>-n] with n = 20, 40, 80 in a fermenter in good yields and confirm their intact protein sequence using various chemical characterization techniques.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70057"},"PeriodicalIF":2.5,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Integrated approach for purification of uricase and protease from Bacillus licheniformis by TPP and IEC.","authors":"Shweta Pawar, Virendra Rathod","doi":"10.1002/btpr.70067","DOIUrl":"https://doi.org/10.1002/btpr.70067","url":null,"abstract":"<p><p>In order to explore the separation and purification methods of uricase and alkaline protease crude enzyme extracts, a combinatorial approach of Three Phase Partitioning (TPP) and Ion-Exchange Chromatography (IEC) was studied. TPP alone was able to separate and purify uricase and alkaline protease enzymes by 5 fold and 2.7 fold, respectively. Further application of ion-exchange chromatography purified enzymes with 99.99% purity in the form of single peaks on a chromatogram. The optimum TPP parameters for simultaneous separation and purification were 50% ammonium sulfate concentration, crude extract to solvent ratio of 1:1, and pH of 8.5. Ion exchange chromatography was performed on a fully automated AKTA start system equipped with a conductivity detector, UV detector, fraction collector, and buffer reservoirs used for isocratic or gradient elution profiles of the proteins under study. Successful purification of enzymes, including their molecular weight, was confirmed with SDS PAGE analysis. Furthermore, the uricase sequence from Bacillus licheniformis was also corroborated to be 87.6% homologous to uricase of B. subtilis using the bioinformatics tool BLASTp (Basic Local Alignment Search Tool for Protein), wherein it compares sequence similarity based on protein or nucleotide sequence. It should be emphasized that this study is the first report for tandem enzyme purification of two enzymes using an automated IEC - AKTA start system where partial purification of enzymes was carried out using TPP.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70067"},"PeriodicalIF":2.5,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940739","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Robert Taylor, Jasdeep Mandur, Umme Amira, Natalie Al-Inati, Juan Marin-Celis, Scott Hatch, Lara Fernandez Cerezo, Nuno Pinto, Efimia Metsi-Guckel, Tiago Matos, Mark Brower, Krunal Mehta, Avik Sarkar
{"title":"Use of live digital twin (shadow) soft sensor to monitor membrane degradation in continuous manufacturing single pass tangential flow filtration.","authors":"Robert Taylor, Jasdeep Mandur, Umme Amira, Natalie Al-Inati, Juan Marin-Celis, Scott Hatch, Lara Fernandez Cerezo, Nuno Pinto, Efimia Metsi-Guckel, Tiago Matos, Mark Brower, Krunal Mehta, Avik Sarkar","doi":"10.1002/btpr.70058","DOIUrl":"https://doi.org/10.1002/btpr.70058","url":null,"abstract":"<p><p>Digital twins (DT) are sophisticated mathematical models representing real-world physical processes, equipped with predictive capabilities that adapt alongside the physical system. The successful implementation of DT in bioprocessing offers numerous advantages, including enhanced understanding of processes, accelerated overall development timelines, and effective monitoring of critical process parameters (CPPs). A comprehensive end-to-end DT can facilitate informed control decisions and forecast how disturbances within the process may affect the final output, accelerating the overall development timelines while optimizing process efficiency and productivity. Tangential flow filtration (TFF) is a standard methodology in bioprocessing, commonly employed to concentrate and exchange buffers for bioproducts. The advancement of continuous process technologies has led to the emergence of alternative TFF methods, notably single-pass tangential flow filtration (SPTFF), which streamlines the process by eliminating the need for stream recirculation. Here, we present the development of a live DT of the SPTFF concentration step within the downstream continuous manufacturing line for a monoclonal antibody (mAb) process. A live DT, equipped with a state estimation tool, was implemented via the Siemens' gPROMS Digital Applications (gDAP) platform. The DT demonstrated the ability to monitor changes in membrane resistance, a typical process parameter that is not directly measured. This parameter is crucial for SPTFF control, as it allows for the constant setting of the concentration factor (CF) by adjusting the retentate flow rate based on the measured resistance and calculated transmembrane pressure (TMP). This achievement illustrates the potential of DT as effective tools for accurately tracking the complete state of the bioprocess.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70058"},"PeriodicalIF":2.5,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144854383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kyeong-Won Yeop, Hyun-Ju Nam, Chang-Jae Shim, So-Mi Yang, Hyo-Won Kim, Cheon Ik Park, Subhasis Banerjee, Yanglin Mok
{"title":"Optimizing clarification processes in biopharmaceutical manufacturing through quality by design: Strategies, implications, and future prospects.","authors":"Kyeong-Won Yeop, Hyun-Ju Nam, Chang-Jae Shim, So-Mi Yang, Hyo-Won Kim, Cheon Ik Park, Subhasis Banerjee, Yanglin Mok","doi":"10.1002/btpr.70063","DOIUrl":"https://doi.org/10.1002/btpr.70063","url":null,"abstract":"<p><p>Biopharmaceutical manufacturing processes in which the product of interest is extracellularly expressed typically employ a clarification step following cell culture or fermentation. During clarification, crude cell culture fluid or fermentation broth is processed to remove insoluble solids, cells, debris, and other particulates, with the extracellular product of interest retained in the filtrate. Soluble impurities, such as host cell proteins (HCPs), may also be partially removed. Historically, the clarification process has been considered a limited contributor to Critical Quality Attributes (CQA). As part of upstream harvest, many biopharmaceutical companies have not fully developed quality control strategies from process development to manufacturing, complicating the application of Quality by Design (QbD) principles to this step. However, advancements in upstream and downstream processing (DSP) technologies, alongside increasing cell counts and titers, necessitate reevaluating clarification as a critical process contributing to drug product quality. Conducting controlled studies to define the process and establish parameters using QbD principles can improve control over process impurities and facilitate a logical quality control strategy, integrating quality into the process. This article describes a systematic approach to QbD for a harvest clarification process where the product of interest is extracellular and impurities are removed in the filtrate post-clarification. It highlights methods for optimizing the clarification unit operation using QbD principles, ensuring better process efficiency, and product quality.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70063"},"PeriodicalIF":2.5,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144844325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Suriyasri Subramanian, Marcin Dembek, Nadia Auchus, Alistair Hines, Paul W A Devine, Åsa Hagner Mcwhirter, Jean-Luc Maloisel, Thomas Linke
{"title":"Enrichment of full AAV capsids by preparative strong anion exchange chromatography.","authors":"Suriyasri Subramanian, Marcin Dembek, Nadia Auchus, Alistair Hines, Paul W A Devine, Åsa Hagner Mcwhirter, Jean-Luc Maloisel, Thomas Linke","doi":"10.1002/btpr.70065","DOIUrl":"https://doi.org/10.1002/btpr.70065","url":null,"abstract":"<p><p>Recombinant adeno-associated virus (rAAV) vectors are the leading in vivo gene delivery platform for the treatment of various human diseases. Scalable manufacturing of rAAV has been successfully demonstrated; however, the presence of non-genome containing empty AAV capsids still remains a significant downstream bottleneck. Separation of empty and full rAAV vectors with linear gradient anion exchange chromatography is challenging to implement at large scale and often achieves only a low recovery of full rAAV capsids. Here we present a workflow to separate empty from full rAAV capsids using Capto Q™ resin with isocratic elution as an alternative. The workflow is based on a preliminary conductivity screening that identifies an optimal empty capsid removal salt concentration, followed by an isocratic two-step elution method. This approach was successfully demonstrated with rAAV serotypes 8 and 9. Approximately 65% of full rAAV8 and rAAV9 capsids were recovered with an enrichment to greater than 80% and 90% full capsids, respectively. Process development using the same approach for rAAV6.2 proved to be more challenging and required a switch in elution salt and an increased concentration of MgCl<sub>2</sub>. The optimized two-step purification protocol for AAV6.2 achieved the recovery of 68% of full capsids with a purity of greater than 80% full capsids.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70065"},"PeriodicalIF":2.5,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144844324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mahmut Alp Kılıç, Mustafa Akyürek, Roozbeh Abidnejad, Alp Karakoç
{"title":"Evaluation of triply periodic minimal surface geometries in 3D-printed PLA scaffolds for chondrogenic differentiation.","authors":"Mahmut Alp Kılıç, Mustafa Akyürek, Roozbeh Abidnejad, Alp Karakoç","doi":"10.1002/btpr.70062","DOIUrl":"https://doi.org/10.1002/btpr.70062","url":null,"abstract":"<p><p>Triply periodic minimal surface (TPMS) scaffolds are gaining attention in tissue engineering due to their continuous and interconnected porous architecture. In this study, three TPMS geometries-Gyroid, Diamond, and I-WP-were fabricated from polylactic acid (PLA) using fused deposition modeling (FDM), with all scaffolds designed to maintain the same overall porosity. Scaffold characterization included scanning electron microscopy (SEM), microcomputed tomography (micro-CT), compressive mechanical testing, and surface wettability analysis. Although porosity was constant, differences in Equivalent Circular Diameter (ECD) values were observed among the geometries, reflecting variations in pore morphology. Adipose-derived stem cells (ADSCs) were seeded onto the scaffolds and cultured under chondrogenic differentiation conditions for 21 days. Cell viability, gene expression (Col2, Col10, Sox9), and protein levels were assessed using RT-PCR and Western blot. All scaffold geometries supported cell attachment and chondrogenic differentiation to varying degrees. The Diamond geometry showed the highest chondrogenic marker expression at the mRNA level, while the Gyroid geometry promoted more stable protein expression with reduced hypertrophic signaling. These findings demonstrate that scaffold geometry, even under identical material and porosity conditions, can influence stem cell behavior. The results offer valuable insights for optimizing TPMS-based scaffold designs in cartilage tissue engineering applications.</p>","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e70062"},"PeriodicalIF":2.5,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144815750","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}