Optimization of rAAV capture step purification using SO3 monolith chromatography.

Adeno-associated virus (AAV) vectors are crucial tools for gene therapy applications. As AAVs are administered in vivo, stringent purity requirements must be met, necessitating the development of various downstream processing strategies in accordance with regulatory guidelines. In this context, we focus on the non-affinity serotype-independent recombinant AAV (rAAV) capture step, which involves the use of Convective Interaction Media (CIM) cation-exchange SO3 monoliths. We analyzed differentially pretreated viral samples obtained from the Sf9 cell line and applied these samples to the capture SO3 chromatography step. We conducted screening experiments using CIM SO3 0.05 mL monolithic 96-well plates with buffers of varying pH, sodium chloride concentrations, and the inclusion of poloxamer 188, aiming to select the optimal binding mobile phase. Dynamic binding capacity was defined for different pretreatments and the optimal conditions were subsequently retested using the industrial purification CIMmultus line. The results demonstrated a high overall vector recovery (51%) and a significant reduction in impurities (99.98% for protein reduction and 99.25% for DNA reduction) using the selected capture step parameters, thereby confirming the successful optimization of the rAAV capture step in the downstream process using monoliths.

Lactic Acid Fermentation of Arthrospira platensis (Spirulina) in a Vegetal Soybean Drink for Developing New Functional Lactose-Free Beverages.

The main objective of this study was to evaluate the suitability of Arthrospira platensis F&M-C256 (spirulina) biomass in a vegetal soybean drink or in water, as substrate for lactic acid fermentation by the probiotic bacterium Lactiplantibacillus plantarum ATCC 8014 (LAB8014) and to evaluate the fermented products in terms of bacteria content and organic acids content, biochemical composition, total phenolics, and phycocyanin content, in vitro digestibility, in vitro and in vivo antioxidant activity. After 72 h of fermentation, a bacterial concentration of about 10.5 log CFU mL-1 in the broths containing the soybean drink + spirulina + LAB8014 (SD + S + LAB8014) or water + spirulina + LAB8014 (W + S + LAB8014) was found. Lactic acid concentration reached similar values (about 1.7 g L-1) in the two broths, while a different acetic acid concentration between SD + S + LAB8014 and W + S + LAB8014 broths was observed (7.7 and 4.1 g L-1, respectively). A. platensis biomass was shown to be a suitable substrate for LAB8014 growth. After fermentation, both broths contained a high protein content (>50%). In both broths, total phenolics, in vitro and in vivo antioxidant activity increased after fermentation (+35, +20, and +93% on average, respectively), while phycocyanin content decreased (-40% on average). Digestibility of W + S + LAB8014 broth statistically improved after fermentation. This study highlights the potential of A. platensis F&M-C256 biomass as a substrate for the production of new functional lactose-free beverages.