Lentiviral Vector Bioprocessing

Lentiviral vectors (LVs) are potent tools for the delivery of genes of interest into mammalian cells and are now commonly utilised within the growing field of cell and gene therapy for the treatment of monogenic diseases and adoptive therapies such as chimeric antigen T-cell (CAR-T) therapy. This is a comprehensive review of the individual bioprocess operations employed in LV production. We highlight the role of envelope proteins in vector design as well as their impact on the bioprocessing of lentiviral vectors. An overview of the current state of these operations provides opportunities for bioprocess discovery and improvement with emphasis on the considerations for optimal and scalable processing of LV during development and clinical production. Upstream culture for LV generation is described with comparisons on the different transfection methods and various bioreactors for suspension and adherent producer cell cultivation. The purification of LV is examined, evaluating different sequences of downstream process operations for both small- and large-scale production requirements. For scalable operations, a key focus is the development in chromatographic purification in addition to an in-depth examination of the application of tangential flow filtration. A summary of vector quantification and characterisation assays is also presented. Finally, the assessment of the whole bioprocess for LV production is discussed to benefit from the broader understanding of potential interactions of the different process options. This review is aimed to assist in the achievement of high quality, high concentration lentiviral vectors from robust and scalable processes

Ultra scale-down approaches to enhance the creation of bioprocesses at scale: impacts of process shear stress and early recovery stages

The sensitivity of biological materials to shear stress conditions encountered during large-scale bioprocessing makes successful scale up from the bench challenging. Ultra scale-down technologies seek to use just millilitre quantities to enhance our understanding of the impact of the process environment as a basis for process optimisation. They can help speed translation of new biological discoveries to market and reduce risks encountered in scale up. They are important both as process discovery tools and as preparative tools to yield material for study of subsequent stages. In this review the focus is on the early recovery stages post fermentation or cell culture and in particular the use of continuous-flow and dead-end centrifugation integrated with preparative stages (e.g. flocculation) and subsequent depth filtration. Examples range from therapeutic antibodies, to rationally engineered (synthetic biology) host strains, to stem cells for therapy

Corak Mazhab Pada Fatwa Dewan Syariah Nasional Majelis Ulama Indonesia (Dsn-Mui)

Penelitian ini menjelaskan mengenai corak mazhab pada fatwa Dewan Syari’ah Nasional MUI terkait tentang Musyarakah, Mudharabah dan Murabahah. Tiga hal yang menjadi fokus utama dalam penelitian ini yakni; 1) kedudukan Mazhab dan Fatwa di Indonesia; 2) eksistensi Dewan Syariah Nasional Majelis Ulama Indonesia, dan; 3) Corak mazhab pada fatwa Dewan Syariah Nasional Majelis Ulama Indonesia mengenai musyarakah, mudharabah dan murabahah. Penelitian ini bertujuan untuk mengetahui corak/tipe mazhab pada fatwa DSN-MUI terkait fatwa mengenai musyarakah, mudharabah dan murabahah. Metode Penelitian yang digunakan oleh penulis dalam penelitian ini adalah menggunakan metode penelitian hukum normatif yaitu metode penelitian hukum terhadap aturan hukum yang tertulis, dimana Fatwa DSN-MUI yang menjadi sumber sekaligus objek penelitian dan sumber data primer dalam penelitian yang dilakukan dan kemudian dianalisis oleh penulis. Berdasakan hasil penelitian maka diperoleh suatu kesimpulan bahwa dalam fatwa DSN-MUI terkait musyarakah, mudharabah dan murabahah, terdapat corak mazhab yang berbeda antar satu sama lain. Pada fatwa DSN-MUI mengenai Musyarakah, jika ditelaah secara substansi, ternyata cenderung bermazhab Hanafi, baik dari segi materi muatan fatwanya maupun secara aplikatif. Lain halnya dengan fatwa DSN-MUI mengenai Mudharabah, fatwa DSN-MUI tersebut lebih banyak mengambil dan menyepakati dasar-dasar hukum yang berasal dari mazhab Syafi’i sebagai fatwanya. Sedangkan fatwa DSN-MUI mengenai Murabahah, dasar hukum dan penetapan substansi dari materi fatwanya, lebih bersandarkan pada proporsi yang sama dari ke empat mazhab yang dikenal selama ini. Dengan demikian, Majelis Ulama Indonesia dalam mengeluarkan fatwa DSN untuk dilaksanakan di Indonesia, selalu mengacu dan berlandaskan dengan mazhab. Meski ada beberapa ketentuan-ketentuan yang disesuaikan dengan kondisi masa dan zaman, tetap tidak menyalahi kaidah-kaidah umum yang telah disampaikan oleh mazhab-mazhab terdahulu

1946 City of Windsor Directory

City of Windsor directory for the year 1946

1949 City of Windsor Directory

City of Windsor directory for the year 1949

1951 City of Windsor Directory

City of Windsor directory for the year 1951

1947 City of Windsor Directory

City of Windsor directory for the year 1947

1945 City of Windsor Directory

City of Windsor directory for the year 1945

1954 City of Windsor Directory

City of Windsor directory for the year 1954

1953 City of Windsor Directory

City of Windsor directory for the year 1953