Razvoj i usporedba alternativnih metoda pročišćavanja adalimumaba izravno iz suspenzije stanične kulture

Research background. Protein A affinity chromatography is a well-established method currently used in the pharmaceutical industry. However, the high costs usually associated with chromatographic separation of protein A and the difficulties in continuous operation make the investigation of alternative purification methods very important. Experimental approach. In this study, extraction/back-extraction and precipitation/dissolution methods were developed and optimised. They were compared with protein A and cation exchange chromatography separations in terms of yield of monoclonal antibody (mAb) and amount of residual impurities, such as DNA and host cell proteins, and amount of mAb aggregates. For a comprehensive comparison of the different methods, experiments were carried out with the same cell-free fermentation broth containing adalimumab. Results and conclusions. Protein A and cation exchange chromatographic separations resulted in high yield and purity of adalimumab. The precipitation-based process resulted in high yield but with lower purity. The extraction-based purification resulted in low yield and purity. Thus, the precipitation-based method proved to be more promising than the extraction-based method for direct purification of adalimumab from harvested cell culture fluid. Novelty and scientific contribution. Although alternative purification methods may offer the advantages of simplicity and low-cost operation, further significant improvements are required to compete with the performance of chromatographic separations of adalimumab from true fermentation broth.Pozadina istraživanja. Afinitetna kromatografija pri kojoj se monoklonsko protutijelo veže za protein A je već dobro poznata metoda u farmaceutskoj industriji. Međutim, zbog visokih troškova izdvajanja proteina A pomoću kromatografije te poteškoća u provođenju kontinuiranog procesa važno je ispitati alternativne metode pročišćavanja. Eksperimentalni pristup. U ovome su radu razvijene i optimirane metode ekstrakcije sa ponovnom ekstrakcijom te taloženja i otapanja. Rezultati tih dvaju metoda, odnosno prinos monoklonskih protutijela, količina preostalih nečistoća, kao što su DNA i proteini stanica domaćina, te količina agregata monoklonskih protutijela, uspoređeni su s onima dobivenim afinitetnom kromatografijom na ionskom izmjenjivaču. Za sveobuhvatnu usporedbu različitih metoda, ispitivanja su provedena na identičnim hranjivim podlogama koje su sadržavale adalimumab. Rezultati i zaključci. Afinitetnom kromatografijom na ionskom izmjenjivaču dobiveni su veliki prinos i velika čistoća adalimumaba. Taloženjem su dobiveni veliki prinos, ali manja čistoća uzorka. Pročišćavanje pomoću ekstrakcije rezultiralo je manjim prinosom i manjom čistoćom uzorka. Zaključeno je da je metoda taloženja bolja od ekstrakcije za izravno pročišćavanje adalimumaba iz suspenzije stanične kulture. Novina i znanstveni doprinos. Iako alternativne metode imaju neke prednosti, kao što su jednostavna i jeftina primjena, neophodno ih je unaprijediti da bi dobiveni rezultati bili usporedivi s rezultatima dobivenim pročišćavanjem adalimumaba ionskom kromatografijom u hranjivoj podlozi

Pharmaceutical and Macromolecular Technologies in the Spirit of Industry 4.0

Well designed (multilayer, adaptive, reactive) interphases, being a key element of multicomponent structures, could be tailored to different requirements through controlled technologies. This is the link connecting various green, safe, healthy materials and innovative pharmaceuticals. Upgraded recycling could be performed by interfacial consolidation of self-reinforced composites, the flame retardancy of which is feasible with surprisingly low amount of flame retardant. Examples are shown how the reinforced and foamed forms of thermosetting and thermoplastic biopolymers can play a significant role (after flame retardant modification) in the development of airplanes and electric cars. Biopolymer nanofibres, such as polycaprolactone and polyhydroxybutyrate, could be formed with increased productivity for various medical uses. Raman-based control of the units of integrated continuous technologies has been elaborated including controlled formation of crystals with polymer interlayer for direct tableting

Oligonucleotide Formulations Prepared by High-Speed Electrospinning: Maximizing Loading and Exploring Downstream Processability

The aim of this study was to develop antisense oligonucleotide tablet formulations using high-speed electrospinning. Hydroxypropyl-beta-cyclodextrin (HPβCD) was used as a stabilizer and as an electrospinning matrix. In order to optimize the morphology of the fibers, electrospinning of various formulations was carried out using water, methanol/water (1:1), and methanol as solvents. The results showed that using methanol could be advantageous due to the lower viscosity threshold for fiber formation enabling higher potential drug loadings by using less excipient. To increase the productivity of electrospinning, high-speed electrospinning technology was utilized and HPβCD fibers containing 9.1% antisense oligonucleotide were prepared at a rate of ~330 g/h. Furthermore, to increase the drug content of the fibers, a formulation with a 50% drug loading was developed. The fibers had excellent grindability but poor flowability. The ground fibrous powder was mixed with excipients to improve its flowability, which enabled the automatic tableting of the mixture by direct compression. The fibrous HPβCD–antisense oligonucleotide formulations showed no sign of physical or chemical degradation over the 1-year stability study, which also shows the suitability of the HPβCD matrix for the formulation of biopharmaceuticals. The obtained results demonstrate possible solutions for the challenges of electrospinning such as scale-up and downstream processing of the fibers

What your 6th grader needs to know : fundamentals of a good sixth-grade education/ Edit.: E.D. Hirsch

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What your 4th grader needs to know : fundamentals of a good fourth-grade education/ Edit.: E.D. Hirsch

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Commodity traders almanac 2012 : for active traders of futures, forex, stocks, and ETFs/ Edit.: Jeffrey A. Hirsch; John L. Person

192 hal.: ill; 23 cm