Pub Date : 2020-01-01DOI: 10.37532/2048-9145.2020.8(1).154-158
H. Li, S. Tian, Z. Liu, J. YuJYan
Objective: To investigate the inhibition of cisplatin-resistant tumor cells on activated CD4+ T cells in ovarian cancer and its mechanism. Methods: 50 patients with ovarian cancer admitted to our hospital from March 2016 to July 2017 were selected as the research group, and 50 normal subjects were selected as the control group. The same separation method of CD4+ T cells was applied to both two groups. The serum levels of ovarian cancer markers and inflammation factors were compared between two groups. Results: The serum levels of ovarian cancer markers in the research group were higher than those in the control group (P<0.05). The average proportion of CD4+ T cells in peasant patients was higher than that in workers and cadres (P<0.05). The average proportion of CD4+ T cells in patients with tumor infiltration was higher than that in patients without infiltration (P<0.05). The level of IL-6 in the research group was higher than that in the control group. While the levels of IL-10, IL-4, IL-2, TNF-α and IFN-γ were lower than those in the control group (P<0.05), and the immune function of the research group was still lower than that of the control group (P<0.05). Conclusion: Cisplatin-resistant ovarian cancer cells can inhibit the activation of CD4+ T cells, which can effectively judge the different stages of ovarian cancer. Moreover, the number of CD4+ T cells can be used to determine the prognosis of patients, providing a certain clue for clinical treatment.
{"title":"Inhibition of cisplatin-resistant tumor cells on activated CD4+ T cells in ovarian cancer and its mechanism","authors":"H. Li, S. Tian, Z. Liu, J. YuJYan","doi":"10.37532/2048-9145.2020.8(1).154-158","DOIUrl":"https://doi.org/10.37532/2048-9145.2020.8(1).154-158","url":null,"abstract":"Objective: To investigate the inhibition of cisplatin-resistant tumor cells on activated CD4+ T cells in ovarian cancer and its mechanism. Methods: 50 patients with ovarian cancer admitted to our hospital from March 2016 to July 2017 were selected as the research group, and 50 normal subjects were selected as the control group. The same separation method of CD4+ T cells was applied to both two groups. The serum levels of ovarian cancer markers and inflammation factors were compared between two groups. Results: The serum levels of ovarian cancer markers in the research group were higher than those in the control group (P<0.05). The average proportion of CD4+ T cells in peasant patients was higher than that in workers and cadres (P<0.05). The average proportion of CD4+ T cells in patients with tumor infiltration was higher than that in patients without infiltration (P<0.05). The level of IL-6 in the research group was higher than that in the control group. While the levels of IL-10, IL-4, IL-2, TNF-α and IFN-γ were lower than those in the control group (P<0.05), and the immune function of the research group was still lower than that of the control group (P<0.05). Conclusion: Cisplatin-resistant ovarian cancer cells can inhibit the activation of CD4+ T cells, which can effectively judge the different stages of ovarian cancer. Moreover, the number of CD4+ T cells can be used to determine the prognosis of patients, providing a certain clue for clinical treatment.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"8 1","pages":"154-158"},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69923705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Y. Kang, R. Ambat, T. Hall, M. D. Sauffer, Stanley, Ng, M. Healy-Fried, Julian Lee, Josaih C Adaelu, W. Holmes, Warren, Emery, Behnam Shanehsaz, A. Huebner, Bo Qi, Richard Chen, Michael, Barry, D. Ludwig, P. Balderes
2015 For ‘acidic’ (pI < 7.0) or ‘neutral’ antibodies (pI: 7.0–8.0), it is challenging to operate traditional anion exchange chromatography in a product flow-through mode to achieve adequate clearance of HCP, DNA, leached ProA, HMW and viruses while maintaining high process yield. In this study, the authors developed a scalable mAb polishing step using a new salt tolerant chromatographic resin. Utilizing a combination of high-throughput condition screening in 96-well plates and optimization in smallscale column models, a polishing step was developed that demonstrated high process yield and efficient clearance of impurities for multiple acidic or neutral antibodies. Pilot scale production demonstrated scalability of the step. This polishing step can be easily integrated into most current Protein A/AEX two-column antibody purification platforms.
{"title":"Development of an acidic / neutral antibody flow-through polishing step using salt-tolerant anion exchange chromatography","authors":"Y. Kang, R. Ambat, T. Hall, M. D. Sauffer, Stanley, Ng, M. Healy-Fried, Julian Lee, Josaih C Adaelu, W. Holmes, Warren, Emery, Behnam Shanehsaz, A. Huebner, Bo Qi, Richard Chen, Michael, Barry, D. Ludwig, P. Balderes","doi":"10.4155/pbp.15.28","DOIUrl":"https://doi.org/10.4155/pbp.15.28","url":null,"abstract":"2015 For ‘acidic’ (pI < 7.0) or ‘neutral’ antibodies (pI: 7.0–8.0), it is challenging to operate traditional anion exchange chromatography in a product flow-through mode to achieve adequate clearance of HCP, DNA, leached ProA, HMW and viruses while maintaining high process yield. In this study, the authors developed a scalable mAb polishing step using a new salt tolerant chromatographic resin. Utilizing a combination of high-throughput condition screening in 96-well plates and optimization in smallscale column models, a polishing step was developed that demonstrated high process yield and efficient clearance of impurities for multiple acidic or neutral antibodies. Pilot scale production demonstrated scalability of the step. This polishing step can be easily integrated into most current Protein A/AEX two-column antibody purification platforms.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/pbp.15.28","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352650","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ricardo J. S. Silva, J. Mota, C. Peixoto, P. Alves, M. Carrondo
Novel biopharmaceutical products, such as vaccines and viral vectors, are a challenging task for downstream processing. Alternative purification strategies that can improve the purification yield, such as continuous chromatography, are regarded nowadays as enabling technologies to overcome the capacity bottleneck in biomanufacturing. This paper reviews the current state of the art in (semi-)continuous chromatographic processes and equipment for purification of bioproducts. Particular focus is given to the recent multicolumn processes developed for virus purification, for which the chromatographic media selection, predictive models, and process design principles are illustrated.
{"title":"Improving the downstream processing of vaccine and gene therapy vectors with continuous chromatography","authors":"Ricardo J. S. Silva, J. Mota, C. Peixoto, P. Alves, M. Carrondo","doi":"10.4155/PBP.15.29","DOIUrl":"https://doi.org/10.4155/PBP.15.29","url":null,"abstract":"Novel biopharmaceutical products, such as vaccines and viral vectors, are a challenging task for downstream processing. Alternative purification strategies that can improve the purification yield, such as continuous chromatography, are regarded nowadays as enabling technologies to overcome the capacity bottleneck in biomanufacturing. This paper reviews the current state of the art in (semi-)continuous chromatographic processes and equipment for purification of bioproducts. Particular focus is given to the recent multicolumn processes developed for virus purification, for which the chromatographic media selection, predictive models, and process design principles are illustrated.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"3 1","pages":"489-505"},"PeriodicalIF":0.0,"publicationDate":"2015-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/PBP.15.29","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The glycosylation profile of a protein is vital to its success as a biotherapeutic as it affects its efficacy and function and, thus, has been extensively studied for proteins produced in Chinese hamster ovary cells. Improvements in the sensitivity and throughput of various analytical assays have allowed for more accurate glycosylation information earlier in the cell line development process. Bioprocess parameters and media additives have been used to modulate the glycosylation of proteins. Genome editing techniques have enabled the regulation of glycosylation genes to achieve a targeted glycan profile. This review highlights advances made in engineering strategies and improvements in high-throughput N-glycan assessment that allow for generation of high-productivity clones with a specific product quality profile.
{"title":"Optimizing Chinese hamster ovary cell line development via targeted control of N-glycosylation","authors":"Christina S. Alves, S. Prajapati","doi":"10.4155/PBP.15.25","DOIUrl":"https://doi.org/10.4155/PBP.15.25","url":null,"abstract":"The glycosylation profile of a protein is vital to its success as a biotherapeutic as it affects its efficacy and function and, thus, has been extensively studied for proteins produced in Chinese hamster ovary cells. Improvements in the sensitivity and throughput of various analytical assays have allowed for more accurate glycosylation information earlier in the cell line development process. Bioprocess parameters and media additives have been used to modulate the glycosylation of proteins. Genome editing techniques have enabled the regulation of glycosylation genes to achieve a targeted glycan profile. This review highlights advances made in engineering strategies and improvements in high-throughput N-glycan assessment that allow for generation of high-productivity clones with a specific product quality profile.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"3 1","pages":"442-461"},"PeriodicalIF":0.0,"publicationDate":"2015-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/PBP.15.25","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hui Cai, M. Guo, A. Gupta, W. Grimm, Andrea Sease, R. Rodriguez, Nesredin A. Mussa, Z. Li
Background: Analysis of process-related protein impurities is critical to ensure process robustness and patient safety. Here, we report on automation of ELISA assays for quantitation of host cell protein and residual protein A. The automated assays were compared with three emerging technologies. Results: Data generated by the automated ELISA platform were comparable to manual results while the throughput was improved by three- to four-times and hands-on time reduced by six- to ten-times. The microfluidic assay enabled the broadest dynamic range and least sample consumption. The bead-based homogeneous assay was the least expensive. The automated ELISA platform demonstrated the highest throughput. Conclusion: Liquid-handler-based automation platform is determined to possess the maximum level of flexibility, adaptability and potential for improvement on assay throughput.
{"title":"Automation of ELISAs & evaluation of emerging technologies for high-throughput quantitation of protein impurities","authors":"Hui Cai, M. Guo, A. Gupta, W. Grimm, Andrea Sease, R. Rodriguez, Nesredin A. Mussa, Z. Li","doi":"10.4155/PBP.15.26","DOIUrl":"https://doi.org/10.4155/PBP.15.26","url":null,"abstract":"Background: Analysis of process-related protein impurities is critical to ensure process robustness and patient safety. Here, we report on automation of ELISA assays for quantitation of host cell protein and residual protein A. The automated assays were compared with three emerging technologies. Results: Data generated by the automated ELISA platform were comparable to manual results while the throughput was improved by three- to four-times and hands-on time reduced by six- to ten-times. The microfluidic assay enabled the broadest dynamic range and least sample consumption. The bead-based homogeneous assay was the least expensive. The automated ELISA platform demonstrated the highest throughput. Conclusion: Liquid-handler-based automation platform is determined to possess the maximum level of flexibility, adaptability and potential for improvement on assay throughput.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"12 1","pages":"427-441"},"PeriodicalIF":0.0,"publicationDate":"2015-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/PBP.15.26","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
J. Brady, Linhong Li, Angelia M. Viley, P. Natarajan, C. Allen, R. Shivakumar, Meg Duskin, M. Peshwa
In the next 5–10 years we could see cellular-based pharmaceuticals, or cell therapy, meeting the unmet medical needs of thousands of people. How this therapy will meet these needs depends on the ability of researchers and manufacturers to successfully and cost effectively manufacture and deliver engineered cell-based therapeutic products that are safe and exhibit enhanced potency with resulting durable, meaningful clinical efficacy. The ability to engineer such enhanced potency using nonviral, cGMP-compliant, automated and closed system manufacturing processes will represent a significant advantage. To outline how such a process might work, we have summarized the application of a scalable, cGMP-compliant, electroporation platform for engineering dendritic cells (DCs), NK cells and T cells for development of cellular immunotherapies targeting hematological malignancies and solid tumors. Autologous cellular immunotherapy refers to a class of therapies that are designed to stimulate a specific immune respons...
{"title":"Development of ‘enhanced’ potency immunotherapy products using nonviral approaches","authors":"J. Brady, Linhong Li, Angelia M. Viley, P. Natarajan, C. Allen, R. Shivakumar, Meg Duskin, M. Peshwa","doi":"10.4155/PBP.15.24","DOIUrl":"https://doi.org/10.4155/PBP.15.24","url":null,"abstract":"In the next 5–10 years we could see cellular-based pharmaceuticals, or cell therapy, meeting the unmet medical needs of thousands of people. How this therapy will meet these needs depends on the ability of researchers and manufacturers to successfully and cost effectively manufacture and deliver engineered cell-based therapeutic products that are safe and exhibit enhanced potency with resulting durable, meaningful clinical efficacy. The ability to engineer such enhanced potency using nonviral, cGMP-compliant, automated and closed system manufacturing processes will represent a significant advantage. To outline how such a process might work, we have summarized the application of a scalable, cGMP-compliant, electroporation platform for engineering dendritic cells (DCs), NK cells and T cells for development of cellular immunotherapies targeting hematological malignancies and solid tumors. Autologous cellular immunotherapy refers to a class of therapies that are designed to stimulate a specific immune respons...","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"3 1","pages":"462-470"},"PeriodicalIF":0.0,"publicationDate":"2015-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/PBP.15.24","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The demand for therapeutic proteins, and particularly monoclonal antibodies, continues to grow within the biopharmaceutical industry. This is illustrated by the monoclonal antibody Humira® (adalimumab) having the highest sales of any drug in 2014. With the increasing numbers of biologics entering the market and the emergence of biosimilars and biobetters, costs need to be reduced in order for companies to remain competitive. High-throughput purification can provide the tools to meet the challenge of producing the thousands of potential drug candidates generated in drug discovery while simultaneously providing robust and cost-effective purification strategies in both research and production. In this review, we discuss the many different methodologies and automation technologies that can be employed in high-throughput purification.
{"title":"Current advances in the development of high-throughput purification strategies for the generation of therapeutic antibodies","authors":"J. Spooner, T. Wilkinson, B. Kemp","doi":"10.4155/PBP.15.23","DOIUrl":"https://doi.org/10.4155/PBP.15.23","url":null,"abstract":"The demand for therapeutic proteins, and particularly monoclonal antibodies, continues to grow within the biopharmaceutical industry. This is illustrated by the monoclonal antibody Humira® (adalimumab) having the highest sales of any drug in 2014. With the increasing numbers of biologics entering the market and the emergence of biosimilars and biobetters, costs need to be reduced in order for companies to remain competitive. High-throughput purification can provide the tools to meet the challenge of producing the thousands of potential drug candidates generated in drug discovery while simultaneously providing robust and cost-effective purification strategies in both research and production. In this review, we discuss the many different methodologies and automation technologies that can be employed in high-throughput purification.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"3 1","pages":"411-424"},"PeriodicalIF":0.0,"publicationDate":"2015-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/PBP.15.23","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In today’s competitive marketplace, the biopharmaceutical industry faces an ever increasing demand to deliver cost-effective therapeutic proteins with limited resources and shrinking timelines. One strategy to increase the throughput of cell culture process development is the utilization of plug-and-play disposable mini-bioreactors (MBRs). These MBRs are capable of operating controlled bioreactor conditions similar to those at traditional bench and large-scale, but in a high-throughput manner without extensive setup or cleanup procedures. These high-throughput systems have been incorporated in various stages of cell culture process development to enable time and resource efficient workflows. This review will highlight the strengths and challenges of several MBR systems within the respective process development workflow and provide insights into the future state of cell culture process development.
{"title":"The role of high-throughput mini- bioreactors in process development and process optimization for mammalian cell culture","authors":"Yao-ming Huang, Chris Kwiatkowski","doi":"10.4155/PBP.15.22","DOIUrl":"https://doi.org/10.4155/PBP.15.22","url":null,"abstract":"In today’s competitive marketplace, the biopharmaceutical industry faces an ever increasing demand to deliver cost-effective therapeutic proteins with limited resources and shrinking timelines. One strategy to increase the throughput of cell culture process development is the utilization of plug-and-play disposable mini-bioreactors (MBRs). These MBRs are capable of operating controlled bioreactor conditions similar to those at traditional bench and large-scale, but in a high-throughput manner without extensive setup or cleanup procedures. These high-throughput systems have been incorporated in various stages of cell culture process development to enable time and resource efficient workflows. This review will highlight the strengths and challenges of several MBR systems within the respective process development workflow and provide insights into the future state of cell culture process development.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"3 1","pages":"397-410"},"PeriodicalIF":0.0,"publicationDate":"2015-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/PBP.15.22","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Generation of recombinant therapeutic proteins involves the use of Chinese hamster ovary cells as one of the workhorses for complex protein production. This process requires screening large numbers of transfected cells to identify single clones that have high protein production and drug-specific quality attributes. Traditionally, this process was limited by manual operation; however, high-throughput screening methods and automation have made this process more efficient. Implementation of high-throughput screening and automation within bioprocess development have led to increased screening capacity, higher product yields, reductions in manual operation, reduction in human error and shorter development time lines. This review outlines the high-throughput methodologies and technologies currently used for clone screening, selection and evaluation in bioprocess development.
{"title":"High-throughput screening and automation approaches for the development of recombinant therapeutic proteins","authors":"Gregory Keil","doi":"10.4155/PBP.15.18","DOIUrl":"https://doi.org/10.4155/PBP.15.18","url":null,"abstract":"Generation of recombinant therapeutic proteins involves the use of Chinese hamster ovary cells as one of the workhorses for complex protein production. This process requires screening large numbers of transfected cells to identify single clones that have high protein production and drug-specific quality attributes. Traditionally, this process was limited by manual operation; however, high-throughput screening methods and automation have made this process more efficient. Implementation of high-throughput screening and automation within bioprocess development have led to increased screening capacity, higher product yields, reductions in manual operation, reduction in human error and shorter development time lines. This review outlines the high-throughput methodologies and technologies currently used for clone screening, selection and evaluation in bioprocess development.","PeriodicalId":90285,"journal":{"name":"Pharmaceutical bioprocessing","volume":"3 1","pages":"371-380"},"PeriodicalIF":0.0,"publicationDate":"2015-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4155/PBP.15.18","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70352345","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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