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  1. Article ; Online: Engineered CHO cells as a novel AAV production platform for gene therapy delivery.

    Nagy, Abdou / Chakrabarti, Lina / Kurasawa, James / Mulagapati, Sri Hari Raju / Devine, Paul / Therres, Jamy / Chen, Zhongying / Schmelzer, Albert E

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 19210

    Abstract: The Herpes simplex virus (HSV)-based platform for production of recombinant adeno-associated viral vectors (rAAVs) yields higher titers and increased percentage of full capsids when compared to the triple transient transfection (TTT) method. However, ... ...

    Abstract The Herpes simplex virus (HSV)-based platform for production of recombinant adeno-associated viral vectors (rAAVs) yields higher titers and increased percentage of full capsids when compared to the triple transient transfection (TTT) method. However, this platform currently faces two major challenges. The first challenge is the reliance on commercial media, sometimes supplemented with serum, leading to costly manufacturing and a high risk for introduction of adventitious agents. The second challenge is that the production of HSV-1 relies on adherent complementing Vero cells (V27), making it difficult to scale up. We engineered serum-free-adapted CHO cells expressing key HSV-1 entry receptors, HVEM and/or Nectin-1 to address the first challenge. Using high-throughput cloning methods, we successfully selected a HVEM receptor-expressing clone (CHO-HV-C1) that yields 1.62 × 10
    MeSH term(s) Cricetinae ; Animals ; Chlorocebus aethiops ; CHO Cells ; Cricetulus ; Vero Cells ; Tissue Distribution ; Herpesvirus 1, Human/genetics ; Genetic Therapy
    Language English
    Publishing date 2023-11-06
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-46298-3
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Accelerated cell culture process development and characterization for cilgavimab/tixagevimab (AZD7442) for the prevention and treatment of COVID-19.

    Handlogten, Michael W / Bosley, Stefanie / Dunn, Sarah / Zhu, Jie / Lee-O'Brien, Allison / Li, Lina / Therres, Jamy / Chakrabarti, Lina / Khanal, Bijay / Mowery, Rachel / Arumugam, Subhashini / Klover, Judith / Taleb, Mohammed / Reier, Jason / Hatton, Diane / Schmelzer, Albert

    Biotechnology and bioengineering

    2023  

    Abstract: The global COVID-19 pandemic ignited an unprecedented race to develop vaccines and antibody therapeutics. AstraZeneca's pursuit to provide AZD7442 (EVUSHELD), two long-acting, SARS-CoV-2 spike receptor binding domain-specific neutralizing monoclonal ... ...

    Abstract The global COVID-19 pandemic ignited an unprecedented race to develop vaccines and antibody therapeutics. AstraZeneca's pursuit to provide AZD7442 (EVUSHELD), two long-acting, SARS-CoV-2 spike receptor binding domain-specific neutralizing monoclonal antibodies, to individuals at risk on highly accelerated timelines challenged our traditional ways of process development and spurred the rapid adoption of novel approaches. Conventional upstream development processes were replaced by agile strategies that combined technological advances and highly accelerated workflows. With calculated business risks and close cross-functional collaborations, this process paved the way for hyper accelerated antibody development from discovery through manufacturing, process validation, emergency use authorization filing, and global regulatory approvals. The result was initiation of commercial manufacturing at a contract manufacturing organization less than 6 months from the selection of cilgavimab and tixagevimab-a process that historically has taken close to 10 years.
    Language English
    Publishing date 2023-02-01
    Publishing country United States
    Document type Journal Article
    ZDB-ID 280318-5
    ISSN 1097-0290 ; 0006-3592
    ISSN (online) 1097-0290
    ISSN 0006-3592
    DOI 10.1002/bit.28336
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.B 960: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
    ab Jg. 2022: Lesesaal (EG)

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  3. Article ; Online: Designing the furin-cleavable linker in recombinant immunotoxins based on Pseudomonas exotoxin A.

    Weldon, John E / Skarzynski, Martin / Therres, Jamy A / Ostovitz, Joshua R / Zhou, Hong / Kreitman, Robert J / Pastan, Ira

    Bioconjugate chemistry

    2015  Volume 26, Issue 6, Page(s) 1120–1128

    Abstract: Recombinant immunotoxins (RITs) are fusion proteins that join antibodies to protein toxins for targeted cell killing. RITs armed with Pseudomonas exotoxin A (PE) are undergoing clinical trials for the treatment of cancer. The current design of PE-based ... ...

    Abstract Recombinant immunotoxins (RITs) are fusion proteins that join antibodies to protein toxins for targeted cell killing. RITs armed with Pseudomonas exotoxin A (PE) are undergoing clinical trials for the treatment of cancer. The current design of PE-based RITs joins an antibody fragment to the catalytic domain of PE using a polypeptide linker that is cleaved by the protease furin. Intracellular cleavage of native PE by furin is required for cytotoxicity, yet the PE cleavage site has been shown to be a poor furin substrate. Here we describe the rational design of more efficiently cleaved furin linkers in PE-based RITs, and experiments evaluating their effects on cleavage and cytotoxicity. We found that changes to the furin site could greatly influence both cleavage and cytotoxicity, but the two parameters were not directly correlated. Furthermore, the effects of alterations to the furin linker were not universal. Identical mutations in the anti-CD22 RIT HA22-LR often displayed different cytotoxicity from mutations in the anti-mesothelin RIT SS1-LR/GGS, underscoring the prominent role of the target site in their intoxication pathways. Combining several beneficial mutations in HA22-LR resulted in a variant (HA22-LR/FUR) with a remarkably enhanced cleavage rate and improved cytotoxicity against five B cell lines and similar or enhanced cytotoxicity in five out of six hairy cell leukemia patient samples. This result informs the design of protease-sensitive linkers and suggests that HA22-LR/FUR may be a candidate for further preclinical development.
    MeSH term(s) ADP Ribose Transferases/chemistry ; ADP Ribose Transferases/metabolism ; ADP Ribose Transferases/pharmacology ; Amino Acid Sequence ; Antineoplastic Agents/chemistry ; Antineoplastic Agents/metabolism ; Antineoplastic Agents/pharmacology ; Bacterial Toxins/chemistry ; Bacterial Toxins/metabolism ; Bacterial Toxins/pharmacology ; Cell Line, Tumor ; Cell Survival/drug effects ; Exotoxins/chemistry ; Exotoxins/metabolism ; Exotoxins/pharmacology ; Furin/metabolism ; Humans ; Immunotoxins/chemistry ; Immunotoxins/metabolism ; Immunotoxins/pharmacology ; Leukemia/drug therapy ; Models, Molecular ; Recombinant Fusion Proteins/chemistry ; Recombinant Fusion Proteins/metabolism ; Recombinant Fusion Proteins/pharmacology ; Virulence Factors/chemistry ; Virulence Factors/metabolism ; Virulence Factors/pharmacology ; Pseudomonas aeruginosa Exotoxin A
    Chemical Substances Antineoplastic Agents ; Bacterial Toxins ; Exotoxins ; Immunotoxins ; Recombinant Fusion Proteins ; Virulence Factors ; ADP Ribose Transferases (EC 2.4.2.-) ; Furin (EC 3.4.21.75)
    Language English
    Publishing date 2015-06-03
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1024041-x
    ISSN 1520-4812 ; 1043-1802
    ISSN (online) 1520-4812
    ISSN 1043-1802
    DOI 10.1021/acs.bioconjchem.5b00190
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 3400: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (2.OG)
    ab Jg. 2022: Lesesaal (EG)

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