Raw material quality assessment approaches comparison in pectin production

Different opportunities are explored to evaluate quality variation in raw materials from biological origin. Assessment of raw materials attributes is an important step in a bio-based production as fluctuations in quality are a major source of process disturbance. This can be due to a variety of biological, seasonal, and supply scarcity reasons. The final properties of a product are invariably linked with the initial properties of the raw material. Thus, the operational conditions of a process can be tuned to drive the product to the required specification based on the quality assessment of the raw material being processed. Process analytical technology tools which enable this assessment in a far more informative and rapid manner than current industrial practices that rely on rule-of-thumb decisions are assessed. An example with citrus peels is used to demonstrate the conceptual and performance differences of distinct quality assessment approaches. The analysis demonstrates the advantage of characterization through multivariate data analysis coupled with a complementary spectroscopic technique, near-infrared spectroscopy. The quantitative comparative analysis of three different approaches, discriminant classification based on expert-knowledge, unsupervised classification, and spectroscopic correlation with reference physicochemical variables, is performed in the same dataset context. © 2018 Her Majesty the Queen in Right of Canada © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2762, 2019.     

Homologous Recombination Offers Advantages over Transposition-Based Systems to Generate Recombinant Baculovirus for Adeno-Associated Viral Vector Production

Viral vectors have a great potential for gene delivery, but manufacturing is a big challenge for the industry. The baculovirus-insect cell is one of the most scalable platforms to produce recombinant adeno-associated virus (rAAV) vectors. The standard procedure to generate recombinant baculovirus is based on Tn7 transposition which is time-consuming and suffers technical constraints. Moreover, baculoviral sequences adjacent to the AAV ITRs are preferentially encapsidated into the rAAV vector particles. This observation raises concerns about safety due to the presence of bacterial and antibiotic resistance coding sequences with a Tn7-mediated system for the construction of baculoviruses reagents. Here, a faster and safer method based on homologous recombination (HR) is investigated. First, the functionality of the inserted cassette and the absence of undesirable genes into HR-derived baculoviral genomes are confirmed. Strikingly, it is found that the exogenous cassette showed increased stability over passages when using the HR system. Finally, both materials generated high rAAV vector genome titers, with the advantage of the HR system being exempted from undesirable bacterial genes which provides an additional level of safety for its manufacturing. Overall, this study highlights the importance of the upstream process and starting biologic materials to generate safer rAAV biotherapeutic products.