Pub Date : 2023-06-01DOI: 10.1016/j.btre.2023.e00791
Gregory M. Jacobson , Kirsty Kraakman , Olivia Wallace , Jolyn Pan , Alex Hennebry , Grant Smolenski , Ray Cursons , Steve Hodgkinson , Adele Williamson , William Kelton
Antigen-specific polyclonal immunoglobulins derived from the serum, colostrum, or milk of immunized ruminant animals have potential as scalable therapeutics for the control of viral diseases including COVID-19. Here we show that the immunization of sheep with fusions of the SARS-CoV-2 receptor binding domain (RBD) to ovine IgG2a Fc domains promotes significantly higher levels of antigen-specific antibodies compared to native RBD or full-length spike antigens. This antibody population contained elevated levels of neutralizing antibodies that suppressed binding between the RBD and hACE2 receptors in vitro. A second immune-stimulating fusion candidate, Granulocyte-macrophage colony-stimulating factor (GM-CSF), induced high neutralizing responses in select animals but narrowly missed achieving significance. We further demonstrated that the antibodies induced by these fusion antigens were transferred into colostrum/milk and possessed cross-neutralizing activity against diverse SARS-CoV-2 variants. Our findings highlight a new pathway for recombinant antigen design in ruminant animals with applications in immune milk production and animal health.
{"title":"Immunogenic fusion proteins induce neutralizing SARS-CoV-2 antibodies in the serum and milk of sheep","authors":"Gregory M. Jacobson , Kirsty Kraakman , Olivia Wallace , Jolyn Pan , Alex Hennebry , Grant Smolenski , Ray Cursons , Steve Hodgkinson , Adele Williamson , William Kelton","doi":"10.1016/j.btre.2023.e00791","DOIUrl":"10.1016/j.btre.2023.e00791","url":null,"abstract":"<div><p>Antigen-specific polyclonal immunoglobulins derived from the serum, colostrum, or milk of immunized ruminant animals have potential as scalable therapeutics for the control of viral diseases including COVID-19. Here we show that the immunization of sheep with fusions of the SARS-CoV-2 receptor binding domain (RBD) to ovine IgG2a Fc domains promotes significantly higher levels of antigen-specific antibodies compared to native RBD or full-length spike antigens. This antibody population contained elevated levels of neutralizing antibodies that suppressed binding between the RBD and hACE2 receptors in vitro. A second immune-stimulating fusion candidate, Granulocyte-macrophage colony-stimulating factor (GM-CSF), induced high neutralizing responses in select animals but narrowly missed achieving significance. We further demonstrated that the antibodies induced by these fusion antigens were transferred into colostrum/milk and possessed cross-neutralizing activity against diverse SARS-CoV-2 variants. Our findings highlight a new pathway for recombinant antigen design in ruminant animals with applications in immune milk production and animal health.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/bc/8f/main.PMC9995299.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9239184","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-06-01DOI: 10.1016/j.btre.2023.e00794
Christine Joy I. Bulaon , Hongyan Sun , Ashwini Malla , Waranyoo Phoolcharoen
The therapeutic blockade of inhibitory PD-1 signaling has emerged as an effective approach for cancer immunotherapy. Nivolumab (Opdivo®), a monoclonal antibody (mAb) targeting the PD-1 immune checkpoint, is approved for treatment of several cancer indications. It functions by blocking the PD-1-mediated T-cell inhibition thus reinstating anticancer immune responses. Tremendous advances in plant biotechnology offer an alternative and economical strategy to produce therapeutic mAbs for immune-based therapies. In this study, recombinant anti-PD-1 Nivolumab was produced in Nicotiana benthamiana and the plant-produced anti-PD-1 mAb was exploited for cancer treatment in syngeneic mice model C57BL/6 mice that were used to test the antitumor efficacy of plant produced Nivolumab, along with commercial Opdivo®. C57BL/6 syngeneic mice treated with plant produced anti-PD-1 mAb exhibited reduction in the growth of established MC38 tumors. The plant produced Nivolumab treatment showed 82.9% antitumor effect in decreasing the tumor volume along with 50% tumor-free mice, whereas Opdivo® showed 90.26% reduction in volume without any tumor-free mice. Finally, plant-derived anti-PD-1 therapy was also well tolerated in tumor-bearing mice that correlated with no significant body weight changes. Overall, our plant-produced Nivolumab elicits significant inhibition of tumor growth in vivo and provides a proof-of-concept for the production of immunotherapy targeting PD-1.
{"title":"Therapeutic efficacy of plant-produced Nivolumab in transgenic C57BL/6-hPD-1 mouse implanted with MC38 colon cancer","authors":"Christine Joy I. Bulaon , Hongyan Sun , Ashwini Malla , Waranyoo Phoolcharoen","doi":"10.1016/j.btre.2023.e00794","DOIUrl":"10.1016/j.btre.2023.e00794","url":null,"abstract":"<div><p>The therapeutic blockade of inhibitory PD-1 signaling has emerged as an effective approach for cancer immunotherapy. Nivolumab (Opdivo®), a monoclonal antibody (mAb) targeting the PD-1 immune checkpoint, is approved for treatment of several cancer indications. It functions by blocking the PD-1-mediated T-cell inhibition thus reinstating anticancer immune responses. Tremendous advances in plant biotechnology offer an alternative and economical strategy to produce therapeutic mAbs for immune-based therapies. In this study, recombinant anti-PD-1 Nivolumab was produced in <em>Nicotiana benthamiana</em> and the plant-produced anti-PD-1 mAb was exploited for cancer treatment in syngeneic mice model C57BL/6 mice that were used to test the antitumor efficacy of plant produced Nivolumab, along with commercial Opdivo®. C57BL/6 syngeneic mice treated with plant produced anti-PD-1 mAb exhibited reduction in the growth of established MC38 tumors. The plant produced Nivolumab treatment showed 82.9% antitumor effect in decreasing the tumor volume along with 50% tumor-free mice, whereas Opdivo® showed 90.26% reduction in volume without any tumor-free mice. Finally, plant-derived anti-PD-1 therapy was also well tolerated in tumor-bearing mice that correlated with no significant body weight changes. Overall, our plant-produced Nivolumab elicits significant inhibition of tumor growth <em>in vivo</em> and provides a proof-of-concept for the production of immunotherapy targeting PD-1.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10090705/pdf/main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9317900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-06-01DOI: 10.1016/j.btre.2023.e00798
Adolfo Henríquez , Juan Pablo Vargas , Chris Landahur , Nicolás Corrales , Andrés Agurto-Muñoz , Pablo A. González , Cristian Agurto-Muñoz
Herpes simplex virus type 2 (HSV-2) is a human infectious agent with significant impact on public health due to its high prevalence in the population and its ability to elicit a wide range of diseases, from mild to severe. Although several antiviral drugs, such as acyclovir, are currently available to treat HSV-2-related clinical manifestations, their effectiveness is poor. Therefore, the identification and development of new antiviral drugs against HSV-2 is necessary. Seaweeds are attractive candidates for such purposes because they are a vast source of natural products due to their highly diverse compounds, many with demonstrated biological activity. In this study, we evaluated the in vitro antiviral potential of red algae extracts obtained from Agarophyton chilense, Mazzaella laminarioides, Porphyridium cruentum, and Porphyridium purpureum against HSV-2. The phycocolloids agar and carrageenan obtained from the macroalgae dry biomass of A. chilense and M. laminarioides and the exopolysaccharides from P. cruentum and P. purpureum were evaluated. The cytotoxicity of these extracts and the surpluses obtained in the extraction process of the agar and carrageenans were evaluated in human epithelial cells (HeLa cells) in addition to their antiviral activity against HSV-2, which were used to calculate selectivity indexes (SIs). Several compounds displayed antiviral activity against HSV-2, but carrageenans were not considered as a potential antiviral therapeutic agent when compared to the other algae extracts with a SI of 23.3. Future assays in vivo models for HSV-2 infection should reveal the therapeutic potential of these algae compounds as new antivirals against this virus.
{"title":"Antiviral activity of red algae phycocolloids against herpes simplex virus type 2 in vitro","authors":"Adolfo Henríquez , Juan Pablo Vargas , Chris Landahur , Nicolás Corrales , Andrés Agurto-Muñoz , Pablo A. González , Cristian Agurto-Muñoz","doi":"10.1016/j.btre.2023.e00798","DOIUrl":"https://doi.org/10.1016/j.btre.2023.e00798","url":null,"abstract":"<div><p>Herpes simplex virus type 2 (HSV-2) is a human infectious agent with significant impact on public health due to its high prevalence in the population and its ability to elicit a wide range of diseases, from mild to severe. Although several antiviral drugs, such as acyclovir, are currently available to treat HSV-2-related clinical manifestations, their effectiveness is poor. Therefore, the identification and development of new antiviral drugs against HSV-2 is necessary. Seaweeds are attractive candidates for such purposes because they are a vast source of natural products due to their highly diverse compounds, many with demonstrated biological activity. In this study, we evaluated the <em>in vitro</em> antiviral potential of red algae extracts obtained from <em>Agarophyton chilense, Mazzaella laminarioides, Porphyridium cruentum,</em> and <em>Porphyridium purpureum</em> against HSV-2. The phycocolloids agar and carrageenan obtained from the macroalgae dry biomass of <em>A. chilense</em> and <em>M. laminarioides</em> and the exopolysaccharides from <em>P. cruentum</em> and <em>P. purpureum</em> were evaluated. The cytotoxicity of these extracts and the surpluses obtained in the extraction process of the agar and carrageenans were evaluated in human epithelial cells (HeLa cells) in addition to their antiviral activity against HSV-2, which were used to calculate selectivity indexes (SIs). Several compounds displayed antiviral activity against HSV-2, but carrageenans were not considered as a potential antiviral therapeutic agent when compared to the other algae extracts with a SI of 23.3. Future assays <em>in vivo</em> models for HSV-2 infection should reveal the therapeutic potential of these algae compounds as new antivirals against this virus.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50176510","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}
Pub Date : 2023-06-01DOI: 10.1016/j.btre.2023.e00793
Drishya M. George , Ramya Ramadoss , Hamish R. Mackey , Annette S. Vincent
{"title":"Corrigendum to Comparative computational study to augment UbiA prenyltransferases inherent in purple photosynthetic bacteria cultured from mangrove microbial mats in Qatar for coenzyme Q10 biosynthesis","authors":"Drishya M. George , Ramya Ramadoss , Hamish R. Mackey , Annette S. Vincent","doi":"10.1016/j.btre.2023.e00793","DOIUrl":"https://doi.org/10.1016/j.btre.2023.e00793","url":null,"abstract":"","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50176573","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}
Pub Date : 2023-06-01DOI: 10.1016/j.btre.2023.e00800
S. Divyashree, B. Shruthi, P.R. Vanitha, M.Y. Sreenivasa
During past twenty years the opportunistic fungal infections have been emerging, causing morbidity and mortality. The fungi belonging to Aspergillus, Mucor, Rhizopus, Candida, Fusarium, Penicillium, Dermatophytes and others cause severe opportunistic fungal infections. Among these Aspergillus and Candida spp cause majority of the diseases. The continuum of fungal infections will prolong to progress in the surroundings of the growing inhabitants of immunocompromised individuals. Presently many chemical-based drugs were used as prophylactic and therapeutic agents. Prolonged usage of antibiotics may lead to some severe effect on the human health. Also, one of the major threats is that the fungal pathogens are becoming the drug resistant. There are many physical, chemical, and mechanical methods to prevent the contamination or to control the disease. Owing to the limitations that are observed in such methods, biological methods are gaining more interest because of the use of natural products which have comparatively less side effects and environment friendly. In recent years, research on the possible use of natural products such as probiotics for clinical use is gaining importance. Probiotics, one of the well studied biological products, are safe upon consumption and are explored to treat various fungal infections. The antifungal potency of major groups of probiotic cultures such as Lactobacillus spp, Leuconostoc spp, Saccharomyces etc. and their metabolic byproducts which act as postbiotics like organic acids, short chain fatty acids, bacteriocin like metabolites, Hydrogen peroxide, cyclic dipeptides etc. to inhibit these opportunistic fungal pathogens have been discussed here.
{"title":"Probiotics and their postbiotics for the control of opportunistic fungal pathogens: A review","authors":"S. Divyashree, B. Shruthi, P.R. Vanitha, M.Y. Sreenivasa","doi":"10.1016/j.btre.2023.e00800","DOIUrl":"10.1016/j.btre.2023.e00800","url":null,"abstract":"<div><p>During past twenty years the opportunistic fungal infections have been emerging, causing morbidity and mortality. The fungi belonging to <em>Aspergillus, Mucor, Rhizopus, Candida, Fusarium, Penicillium, Dermatophytes</em> and others cause severe opportunistic fungal infections. Among these <em>Aspergillus</em> and <em>Candida spp</em> cause majority of the diseases. The continuum of fungal infections will prolong to progress in the surroundings of the growing inhabitants of immunocompromised individuals. Presently many chemical-based drugs were used as prophylactic and therapeutic agents. Prolonged usage of antibiotics may lead to some severe effect on the human health. Also, one of the major threats is that the fungal pathogens are becoming the drug resistant. There are many physical, chemical, and mechanical methods to prevent the contamination or to control the disease. Owing to the limitations that are observed in such methods, biological methods are gaining more interest because of the use of natural products which have comparatively less side effects and environment friendly. In recent years, research on the possible use of natural products such as probiotics for clinical use is gaining importance. Probiotics, one of the well studied biological products, are safe upon consumption and are explored to treat various fungal infections. The antifungal potency of major groups of probiotic cultures such as <em>Lactobacillus spp, Leuconostoc spp, Saccharomyces</em> etc. and their metabolic byproducts which act as postbiotics like organic acids, short chain fatty acids, bacteriocin like metabolites, Hydrogen peroxide, cyclic dipeptides etc. to inhibit these opportunistic fungal pathogens have been discussed here.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/49/ba/main.PMC10196798.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9859390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-06-01DOI: 10.1016/j.btre.2023.e00801
Paulina Korpys-Woźniak, Ewelina Celińska
While the unfolded protein response (UPR) and its major regulator – transcription factor Hac1 are well-conserved across Eukarya, species-specific variations are repeatedly reported. Here we investigated molecular mechanisms by which co-over-expression of HAC1 improves secretion of a recombinant protein (r-Prot) in Yarrowia lipolytica, using comparative transcriptomics. Co-over-expression of HAC1 caused an >2-fold increase in secreted r-Prot, but its intracellular levels were decreased. The unconventional splicing rate of the HAC1 mRNA was counted through transcript sequencing. Multiple biological processes were affected in the HAC1-and-r-Prot co-over-expressing strain, including ribosome biogenesis, nuclear and mitochondrial events, cell cycle arrest, attenuation of gene expression by RNA polymerase III and II, as well as modulation of proteolysis and RNA metabolism; but whether the HAC1 co-over-expression/induction was the actual causative agent for these changes, was not always clear. We settled that the expression of the “conventional” HAC1 targets (KAR2 and PDI1) is not affected by its over-expression.
{"title":"Molecular background of HAC1-driven improvement in the secretion of recombinant protein in Yarrowia lipolytica based on comparative transcriptomics","authors":"Paulina Korpys-Woźniak, Ewelina Celińska","doi":"10.1016/j.btre.2023.e00801","DOIUrl":"https://doi.org/10.1016/j.btre.2023.e00801","url":null,"abstract":"<div><p>While the unfolded protein response (UPR) and its major regulator – transcription factor Hac1 are well-conserved across <em>Eukarya</em>, species-specific variations are repeatedly reported. Here we investigated molecular mechanisms by which co-over-expression of <em>HAC1</em> improves secretion of a recombinant protein (r-Prot) in <em>Yarrowia lipolytica</em>, using comparative transcriptomics. Co-over-expression of <em>HAC1</em> caused an >2-fold increase in secreted r-Prot, but its intracellular levels were decreased. The unconventional splicing rate of the <em>HAC1</em> mRNA was counted through transcript sequencing. Multiple biological processes were affected in the <em>HAC1</em>-and-r-Prot co-over-expressing strain, including ribosome biogenesis, nuclear and mitochondrial events, cell cycle arrest, attenuation of gene expression by RNA polymerase III and II, as well as modulation of proteolysis and RNA metabolism; but whether the <em>HAC1</em> co-over-expression/induction was the actual causative agent for these changes, was not always clear. We settled that the expression of the “conventional” <em>HAC1</em> targets (<em>KAR2</em> and <em>PDI1</em>) is not affected by its over-expression.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50176511","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}
Pub Date : 2023-06-01DOI: 10.1016/j.btre.2023.e00799
Aya Abdel-Nasser , Amal S. Hathout , Ahmed N. Badr , Olfat S. Barakat , Hayam M. Fathy
Aflatoxins are toxic carcinogens and mutagens formed by some moulds, specifically Aspergillus spp. Therefore, this study aimed to extract and identify bioactive secondary metabolites from Lactobacillus species, to evaluate their efficacy in reducing fungal growth and aflatoxin production and to investigate their toxicity. The bioactive secondary metabolites of Lactobacillus species showed variable degrees of antifungal activity, whereas L. rhamnosus ethyl acetate extract No. 5 exhibited the highest antifungal activity and, thus, was selected for further identification studies. Data revealed that L. rhamnosus ethyl acetate extract No. 5 produced various organic acids, volatile organic compounds and polyphenols, displayed antifungal activity against A. flavus, and triggered morphological changes in fungal conidiophores and conidiospores. L. rhamnosus ethyl acetate extract No. 5 at a 9 mg/mL concentration reduced AFB1 production by 99.98%. When the effect of L. rhamnosus ethyl acetate extract No. 5 on brine shrimp mortality was studied, the extract attained a 100% mortality at a concentration of 400 µg/mL, with an IC50 of 230 µg/mL. Meanwhile, a mouse bioassay was performed to assess the toxicity of L. rhamnosus ethyl acetate extract No. 5, whereas there were no harmful effects or symptoms in mice injected with L. rhamnosus ethyl acetate extract at concentrations of 1, 3, 5, 7, and 9 mg/kg body weight.
{"title":"Extraction and characterization of bioactive secondary metabolites from lactic acid bacteria and evaluating their antifungal and antiaflatoxigenic activity","authors":"Aya Abdel-Nasser , Amal S. Hathout , Ahmed N. Badr , Olfat S. Barakat , Hayam M. Fathy","doi":"10.1016/j.btre.2023.e00799","DOIUrl":"10.1016/j.btre.2023.e00799","url":null,"abstract":"<div><p>Aflatoxins are toxic carcinogens and mutagens formed by some moulds, specifically <em>Aspergillus</em> spp. Therefore, this study aimed to extract and identify bioactive secondary metabolites from <em>Lactobacillus</em> species, to evaluate their efficacy in reducing fungal growth and aflatoxin production and to investigate their toxicity. The bioactive secondary metabolites of <em>Lactobacillus</em> species showed variable degrees of antifungal activity, whereas <em>L. rhamnosus</em> ethyl acetate extract No. 5 exhibited the highest antifungal activity and, thus, was selected for further identification studies. Data revealed that <em>L. rhamnosus</em> ethyl acetate extract No. 5 produced various organic acids, volatile organic compounds and polyphenols, displayed antifungal activity against <em>A. flavus</em>, and triggered morphological changes in fungal conidiophores and conidiospores. <em>L. rhamnosus</em> ethyl acetate extract No. 5 at a 9 mg/mL concentration reduced AFB<sub>1</sub> production by 99.98%. When the effect of <em>L. rhamnosus</em> ethyl acetate extract No. 5 on brine shrimp mortality was studied, the extract attained a 100% mortality at a concentration of 400 µg/mL, with an IC<sub>50</sub> of 230 µg/mL. Meanwhile, a mouse bioassay was performed to assess the toxicity of <em>L. rhamnosus</em> ethyl acetate extract No. 5, whereas there were no harmful effects or symptoms in mice injected with <em>L. rhamnosus</em> ethyl acetate extract at concentrations of 1, 3, 5, 7, and 9 mg/kg body weight.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/4a/b8/main.PMC10189384.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9551563","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Human induced pluripotent stem cells (iPSC) have demonstrated massive potentials for use in regenerative and personalized medicine due to their ability to expand in culture and differentiate into specialized cells with therapeutic benefits. However, in order to industrialize iPSC-derived therapies, it is necessary to address the existing challenges surrounding the analytics implemented in the manufacturing process to evaluate and monitor cell expansion, differentiation, and quality of the final products. Here, we review some of the key analytical methods used as part of identity, potency, or safety for in-process or final product release testing and highlighted the challenges and potential solutions for consideration in the Chemistry, Manufacturing and Controls (CMC) strategy for iPSC-based therapies.
Some of the challenges associated with characterization and testing of iPSC-based products are related to the choice of analytical technology (to ensure fit-for-purpose), assay reliability and robustness. Automation of analytical methods may be required to reduce hands on time, and improve reliability of the methods through reducing assay variability. Indeed, we have shown that automation of analytical methods is feasible (evaluated using an ELISA based assay) and would result in more precise measurements (demonstrated by lower co-efficient of Variation and standard deviation), less hands-on time, and swift compared to a manually run assay. Therefore, in order to support commercialization of iPSC-based therapies we suggest a well-designed testing strategy to be established in the development phase while incorporating robust, reproducible, reliable, and potentially automated analytics in the manufacturing process.
{"title":"Characterization of human induced pluripotent stems cells: Current approaches, challenges, and future solutions","authors":"Sahana Suresh Babu, Haritha Duvvuru, Jillian Baker, Stephanie Switalski, Mehdi Shafa, Krishna Morgan Panchalingam, Saedeh Dadgar, Justin Beller, Behnam Ahmadian Baghbaderani","doi":"10.1016/j.btre.2023.e00784","DOIUrl":"10.1016/j.btre.2023.e00784","url":null,"abstract":"<div><p>Human induced pluripotent stem cells (iPSC) have demonstrated massive potentials for use in regenerative and personalized medicine due to their ability to expand in culture and differentiate into specialized cells with therapeutic benefits. However, in order to industrialize iPSC-derived therapies, it is necessary to address the existing challenges surrounding the analytics implemented in the manufacturing process to evaluate and monitor cell expansion, differentiation, and quality of the final products. Here, we review some of the key analytical methods used as part of identity, potency, or safety for in-process or final product release testing and highlighted the challenges and potential solutions for consideration in the Chemistry, Manufacturing and Controls (CMC) strategy for iPSC-based therapies.</p><p>Some of the challenges associated with characterization and testing of iPSC-based products are related to the choice of analytical technology (to ensure fit-for-purpose), assay reliability and robustness. Automation of analytical methods may be required to reduce hands on time, and improve reliability of the methods through reducing assay variability. Indeed, we have shown that automation of analytical methods is feasible (evaluated using an ELISA based assay) and would result in more precise measurements (demonstrated by lower co-efficient of Variation and standard deviation), less hands-on time, and swift compared to a manually run assay. Therefore, in order to support commercialization of iPSC-based therapies we suggest a well-designed testing strategy to be established in the development phase while incorporating robust, reproducible, reliable, and potentially automated analytics in the manufacturing process.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/aa/c0/main.PMC9929203.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10767425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Marine macroalgae are being recognized as reservoirs of biologically active compounds, as their surfaces are susceptible to the colonization of microorganisms which can produce enzymes with a wide range of molecular architectures. Among these bacteria, Achromobacter is responsible for the biosynthesis of laccases. In this research, we performed a bioinformatic pipeline to annotate the sequenced complete genome of the epiphytic bacterium Achromobacter denitrificans strain EPI24, from the macroalgal surface of the Ulva lactuca species; this strain showed laccase activity which has been previously assessed on plate assays. The genome of A. denitrificans strain EPI24 has a size of ∼6.95 Mb, a GC content of 67.33%, and 6,603 protein-coding genes. The functional annotation of the A. denitrificans strain EPI24 genome confirmed the presence of genes encoding for laccases, which could have functional properties of interest in processes such as the biodegradation of phenolic compounds under versatile and efficient conditions.
{"title":"Genome of the epiphytic bacterium Achromobacter denitrificans strain EPI24, isolated from a macroalga located in the Colombian Caribbean","authors":"Albert Nicolás Niño Corredor , Natalia Beatriz Comba González , Mauricio Acelas , Dolly Montoya Castaño","doi":"10.1016/j.btre.2023.e00788","DOIUrl":"https://doi.org/10.1016/j.btre.2023.e00788","url":null,"abstract":"<div><p>Marine macroalgae are being recognized as reservoirs of biologically active compounds, as their surfaces are susceptible to the colonization of microorganisms which can produce enzymes with a wide range of molecular architectures. Among these bacteria, <em>Achromobacter</em> is responsible for the biosynthesis of laccases. In this research, we performed a bioinformatic pipeline to annotate the sequenced complete genome of the epiphytic bacterium <em>Achromobacter denitrificans</em> strain EPI24, from the macroalgal surface of the <em>Ulva lactuca</em> species; this strain showed laccase activity which has been previously assessed on plate assays. The genome of <em>A. denitrificans</em> strain EPI24 has a size of ∼6.95 Mb, a GC content of 67.33%, and 6,603 protein-coding genes. The functional annotation of the <em>A. denitrificans</em> strain EPI24 genome confirmed the presence of genes encoding for laccases, which could have functional properties of interest in processes such as the biodegradation of phenolic compounds under versatile and efficient conditions.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50182348","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}
This work aimed to carry out chemical cooking of corn stalks, both in a nitrate-alkaline manner and in a soda pulp method. The composition of corn is characterized by cellulose, lignin, ash, and substances extractable into polar and organic solvents. Handsheets were made from the pulp, for which the degree of polymerization, sedimentation rate, and strength properties was determined.
{"title":"Chemical pulp from corn stalks","authors":"Kateřina Hájková , Tereza Jurczyková , Michaela Filipi , Jiří Bouček","doi":"10.1016/j.btre.2023.e00786","DOIUrl":"10.1016/j.btre.2023.e00786","url":null,"abstract":"<div><p>This work aimed to carry out chemical cooking of corn stalks, both in a nitrate-alkaline manner and in a soda pulp method. The composition of corn is characterized by cellulose, lignin, ash, and substances extractable into polar and organic solvents. Handsheets were made from the pulp, for which the degree of polymerization, sedimentation rate, and strength properties was determined.</p></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/d7/e3/main.PMC9945710.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10792428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}