Purification of rAAV is a crucial unit operation of the AAV production process. It enables the capture of AAV and removal of contaminants such as host cell proteins, host cell DNA, and other cell culture-related impurities. Here we describe the purification of rAAV produced in insect cells Sf9/rBEV by immuno-affinity capture chromatography. The method is fully scale-amenable unlike other traditional purification methods based on ultracentrifugation. The method reported herein has two main steps: (1) the clarification of cell lysate by depth filtration and (2) the selective capture and single-step purification of AAV via immune-affinity chromatography. This purification method has been successfully implemented to purify the majority of wild-type AAV serotypes.

The clarification conditions and the selection of the clarification agent are pivotal in eliminating the haze components from red grape juice (RGJ) while minimizing the loss of functional color components. In this context, we synthesized a water glass-based APTES functionalized magnesium silica aerogel (MSA-NH3) incorporating 61.44 molecules/nm2 of amine groups, resulting in a positively charged zeta potential value of 33.9 mV (pH 3.4) for clarification of RGJ by targeting negatively charged polyphenols. The optimum clarification conditions using MSA-NH3 were determined as 0.18 g MSA-NH3/L RGJ, 20 °C, and 60 min through the application of Box-Behnken design. Under these conditions, MSA-NH3 exhibited excellent adsorption of haze components (3.61 NTU), outperforming the commercial bentonite-gelatine combination (BGC) (5.45 NTU). Furthermore, it exhibited greater efficacy in preserving anthocyanins while adsorbing browning components. MSA-NH3 has a high potential to serve as a functional alternative clarification agent in the beverage industry due to its promising clarification performance.

This study aimed at using α-L-arabinofuranosidase CcABF to improve the clarity and active substances in fermented ginkgo kernel juice by artificial neural network (ANN) modeling and genetic algorithm (GA) optimization. A credible three-layer feedforward ANN model was established to predict the optimal parameters for CcABF clarification. The experiments proved the highest transmittance of 89.40% for fermented ginkgo kernel juice with this understanding, which exhibited a 25.56% increase over the unclarified group. With the clarification of CcABF, the antioxidant capacity in juice was enhanced with the increase of total phenolic and flavone contents, and the maximum DPPH and hydroxyl radical scavenging rates were increased by 89.71% and 26.65%, respectively. The contents of toxic ginkgolic acids declined markedly, while the active ingredients of ginkgetin and ginkgolide B showed a modest increase. Moreover, changes in free amino acids and volatile compounds improved the nutritive value and flavor of clarified fermented ginkgo kernel juice.

The use of ophthalmic agents during pregnancy and breastfeeding always represents an off-label use. Therefore, the use of drugs must be particularly carefully assessed with respect to the risk-benefit assessment. In this overview the literature databank of the PubMed library, pharmaceutical lists (Red List, Swiss pharmaceutical compendium), guidelines of the specialist societies the German Society of Ophthalmology (DOG), the Swiss Society of Ophthalmology (SOG), the European Glaucoma Society (EGS), the American Academy of Ophthalmology (AAO) and internet portals (embryotox, reprotox) were inspected and recommendations for the use of ophthalmic agents during pregnancy and breastfeeding were derived. More attention should be dedicated to this topic in the specialist societies.
UNASSIGNED: Die Verwendung von Ophthalmika in der Schwangerschaft und Stillzeit stellt immer eine Off-label-Anwendung dar. Ein Einsatz von Arzneimitteln muss daher besonders sorgfältig auf sein Risiko-Nutzen-Verhältnis bewertet werden. In der vorliegenden Übersicht wurden Literaturverzeichnisse (Pubmed Library), Arzneimittelverzeichnisse (Rote Liste, Schweizer Arzneimittelkompendium), Leitlinien von Fachgesellschaften (Deutsche Ophthalmologische Gesellschaft [DOG], Schweizerische Ophthalmologische Gesellschaft [SOG], European Glaucoma Society [EGS], American Academy of Ophthalmology [AAO]) und Internetportale (Embryotox, Reprotox) gesichtet und daraus Empfehlungen für den Einsatz von Ophthalmika in der Schwangerschaft und Stillzeit abgeleitet. Dem Thema sollte mehr Aufmerksamkeit in den Fachgesellschaften gewidmet werden.

The Orf virus (ORFV) is a promising vector platform for the generation of vaccines against infectious diseases and cancer, highlighted by its progression to clinical testing phases. One of the critical steps during GMP manufacturing is the clarification of crude harvest because of the enveloped nature and large size of ORFV. This study presents the first description of ORFV clarification process from a HEK suspension batch process. We examined various filter materials, membrane pore sizes, harvest timings, and nuclease treatments. Employing the Ambr® crossflow system for high-throughput, small-volume experiments, we identified polypropylene-based Sartopure® PP3 filters as ideal. These filters, used in two consecutive stages with reducing pore sizes, significantly enhanced ORFV recovery and addressed scalability challenges. Moreover, we demonstrated that the time of harvest and the use of a nuclease play a decisive role to increase ORFV yields. With these findings, we were able to establish an efficient and scalable clarification process of ORFV derived from a suspension production process, essential for advancing ORFV vaccine manufacturing.

暂无摘要。

Using clarifying agents is essential in the production of fruit juice. This study utilized gelatin and bentonite as clarifying agents to improve the quality and shelf-life of Indian gooseberry juice (IGBJ). Different treatments were prepared using varying levels of gelatin and bentonite alone or in combination (1.5-2 mg/mL and 1-2 mg/mL, respectively). The untreated IGBJ was used as a control. The results showed the combined treatment of 1.5 mg/mL gelatin and 1 mg/mL bentonite significantly (p ≤ 0.05) improved the transmittance, △E value, total acidity, vitamin C, and antioxidant activity of the IGBJ sample. During storage for 49 days at 4 °C, the quality changes in the IGBJ were minimal with the use of 1.5 mg/mL gelatin and 1 mg/mL bentonite compared to the control (p > 0.05). The treated samples showed no signs of spoilage bacteria, yeast, or mold during cold storage. The combined use of gelatin and bentonite (1.5 mg/mL gelatin and 1 mg/mL bentonite) was found to effectively preserve the sensory quality, bioactivity, and color properties of IGBJ, thereby extending its shelf-life. Thus, gelatin and bentonite can be used as preferred filtering aids for quality and shelf-life extension in the food industry, as they have synergistic effects.

The recovery of valuable compounds like phenolic compounds and sugars from grape marc extracts implies different steps, including clarification. In this study, a response surface methodology (RSM) was used as a statistical tool to study the effects of operating conditions such as transmembrane pressure (TMP), temperature and feed flow rate on the performance of a microfiltration (MF) monotubular ceramic membrane with a pore size of 0.14 μm in the clarification of grape marc extract from the Carménère variety, as well to optimize the process conditions by implementing the Box-Behnken statistical design. The desirability function approach was applied to analyze the regression model equations in order to maximize the permeate flux and concentration of malvidin-3-O-glucoside, glucose and fructose in the clarified extract. The optimal operating conditions were found to be 1 bar, 29.01 °C and 5.64 L/min. Under these conditions, the permeate flux and concentration of malvidin-3-O-glucoside, glucose and fructose resulted in 65.78 L/m2h, 43.73 mg/L, 305.89 mg/L, and 274.85 mg/L, respectively.

Protein A capture chromatography remains a high-cost and relatively low-productivity step for downstream processing of monoclonal antibodies. As bioprocessing transitions toward intensified processes, maximizing the efficiency of individual steps is key to achieving economic targets. This study was performed to assess the impact of inline concentration of clarified cell culture fluid (CCF), using single-pass tangential flow filtration, on protein A chromatography purification productivity. CCF with varying levels of impurities and turbidity were obtained dependent upon the clarification method. These CCFs were concentrated and processed over a protein A capture step. Productivity increases of 1.8- to 2.6-fold were achieved as compared to a protein A capture step with no CCF concentration. Achieving such targeted improvements requires careful consideration of the multiple components in the clarification strategy before implementation.

BACKGROUND: Flavor is considered as a key quality attribute of fruit juice affecting consumer acceptance. During processing, the flavor loss of cloudy juice always occurs due to the variations of juice cloud particles. Pectin, a major component of cloud particles, plays an important role in cloud stability. In this work, we focused on the effects of variation of three pectin fractions caused by gentle centrifugation and clarification on the physicochemical properties, volatile content and sensory profile of heat-sterilized muskmelon cloudy juice.
RESULTS: Centrifugation treatment reduced the total soluble solids and viscosity of cloudy juice and increased cloud stability. With centrifugation increased, the contents of most monosaccharides in the three pectin fractions were reduced. Most aroma-active aldehydes and alcohols, such as (2E,6Z)-nonadienal, 1-octen-3-ol and (E)-non-2-enal, after gentle centrifugation and clarification, were maintained, but most esters were decreased. The volatile compositions were highly related to the three pectin fractions. The addition of chelator-soluble pectin and sodium carbonate-soluble pectin could decrease the formation of dimethyl trisulfide and dimethyl disulfide in clarified juice, thereby improving the sensory profile.
CONCLUSIONS: The results suggested that endogenous chelator-soluble pectin and sodium carbonate-soluble pectin can be used in heat-sterilized fruit juice to improve flavor quality, with an emphasis on a significant reduction in volatile sulfur compounds. © 2023 Society of Chemical Industry.