Der deutsche Dichter Heinrich Heine (1797–1856) zählt zu den herausragendsten und gleichzeitig kontroversesten Persönlichkeiten seiner Zeit, welche später in den deutschen literarischen Kanon ...aufgenommen wurden. Die Rezeption Heines in der Vergangenheit zeichnet sich durch wiederholte Ablehnung aus, da sowohl die Gesellschaft als auch Kritiker seinen politischen und religiösen Ansichten eher negativ gegenüberstanden. Der vorliegende Beitrag konzentriert sich auf die Rezeption Heinrich Heines im slowenischen ethnischen Gebiet bis zum Jahr 1860 und gliedert sich in drei Kapitel. Das erste Kapitel untersucht Heines Rezeption vor dem Jahr 1856, das zweite Kapitel beleuchtet seine Rezeption im Todesjahr des Dichters 1856, während sich das dritte Kapitel der Rezeption zwischen den Jahren 1857 und 1860 widmet. Die Studie umfasst eine Analyse von Veröffentlichungen in insgesamt neun journalistischen Organen, die zu dieser Zeit auf dem slowenischen ethnischen Gebiet herausgegeben wurden. Die untersuchten Zeitungen sind die Laibacher Zeitung samt ihren Beilagen, dem Intelligenzblatt zur Laibacher Zeitung, dem Illyrischen Blatt und den Blättern aus Krain, die Carinthia, die Carniolia, die Kmetijske in rokodelske novice, die Zgodnja danica sowie die literarische Zeitschrift Slovenski glasnik. Heines Präsenz ist bis zum Jahr 1860 in nahezu allen genannten Zeitungen mindestens einmal nachweisbar, mit Ausnahme der literarischen Zeitschrift Slovenski glasnik.
A method to separate VLP from extra cellular vesicles.Extra cellular particles have similar size range as VLPs.After separation detected by proteomics tools.Method in principle suited for ...manufacturing of pandemic vaccine.
Enveloped virus-like particles (VLPs) are increasingly used as vaccines and immunotherapeutics. Frequently, very time consuming density gradient centrifugation techniques are used for purification of VLPs. However, the progress towards optimized large-scale VLP production increased the demand for fast, cost efficient and scale able purification processes. We developed a chromatographic procedure for purification of HIV-1 gag VLPs produced in CHO cells. The clarified and filtered cell culture supernatant was directly processed on an anion-exchange monolith. The majority of host cell impurities passed through the column, whereas the VLPs were eluted by a linear or step salt gradient; the major fraction of DNA was eluted prior to VLPs and particles in the range of 100⿿200nm in diameter could be separated into two fractions. The earlier eluted fraction was enriched with extracellular particles associated to exosomes or microvesicles, whereas the late eluting fractions contained the majority of most pure HIV-1 gag VLPs. DNA content in the exosome-containing fraction could not be reduced by Benzonase treatment which indicated that the DNA was encapsulated. Many exosome markers were identified by proteomic analysis in this fraction. We present a laboratory method that could serve as a basis for rapid downstream processing of enveloped VLPs. Up to 2000 doses, each containing 1ÿ109 particles, could be processed with a 1mL monolith within 47min. The method compared to density gradient centrifugation has a 220-fold improvement in productivity.
The NanoSight LM10 with Nanoparticle tracking analysis (NTA) software was evaluated for the quantification of latex particles, adenovirus 5, and influenza virus. The inter-day variability was ...determined by measuring the same sample over several consecutive days and the method's accuracy was demonstrated by using known concentrations of the subject particles. NTA analysis was also used to quantify chromatographic fractions of adenovirus and influenza virus after purification on a CIM monolithic column. NTA results were compared and evaluated against hemagglutination (HA) and end point dilution assay, determining total and infection virus particle number, respectively. The results demonstrated that nanoparticle tracking analysis is a method for fast estimation of virus concentration in different samples. In addition, it can provide a better insight into the sample status, regarding the level of virus aggregation.
The downstream processing of enveloped virus‐like particles is very challenging because of the biophysical and structural similarity between correctly assembled particles and contaminating vesicular ...particles present in the feedstock. We used hydroxyl‐functionalized polymethacrylate monoliths, providing hydrophobic and electrostatic binding contributions, for the purification of HIV‐1 gag virus‐like particles. The clarified culture supernatant was conditioned with ammonium sulfate and after membrane filtration loaded onto a 1 mL monolith. The binding capacity was 2 × 1012/mL monolith and was only limited by the pressure drop. By applying either a linear or a step gradient elution, to decrease the ammonium sulfate concentration, the majority of double‐stranded DNA (88–90%) and host cell protein impurities (39–61%) could be removed while the particles could be separated into two fractions. Proteomic analysis and evaluation of the p24 concentration showed that one fraction contained majority of the HIV‐1 gag and the other fraction was less contaminated with proteins originated from intracellular compartments. We were able to process up to 92 bed volumes of conditioned loading material within 3 h and eluted in average 7.3 × 1011 particles per particle fraction, which is equivalent to 730 vaccination doses of 1 × 109 particles.
•Direct chromatographic processing of virus without pre-concentration.•Protocol for rapid purification of infective virus.•Purification method to yield >90% infective virus.•Phospholipid removal ...prior anion-exchange chromatography essential for recovery of virus.
A chromatographic process based on monoliths for purification of infective baculovirus without prior concentration step has been established. Baculovirus produced in Spodoptera frugiperda cells (Sf-9) were harvested by centrifugation, filtered through 0.8μm filters and directly loaded onto radial 1mL anion exchange monoliths with a channel size of 1.5–2.0μm operated at a volumetric flow rate of one bed volume per minute. Optional an epoxy monolith was used as pre-column to reduce interfering compounds and substances influencing the capacity of anion exchange monoliths for baculovirus infectious virus could be eluted with a step gradient at salt concentrations of 440mM NaCl. Recovery of infectious virus was highly influenced by composition and age of supernatant and ranged from 20 to >99% active baculovirus. Total protein content could be reduced to 1–8% and DNA content to 38–48% in main virus fraction. Infective virus could be 52-fold concentrated within 20.5h and simultaneously an 82-fold volume reduction was possible when loading 1150mL (2.1×108pfu/mL) onto 1mL scale support.
Highlights • We described novel chromatographic influenza virus vaccine purification method. • Method is robust enabling purification of different influenza A and B subtypes. • Method yields up to ...100% infectious virus. • High vaccine purity was achieved with >99% host cell DNA removal and >95% protein removal. • Method was used for production of attenuated, replication deficient influenza vaccine for clinical trials.
Der deutsche Dichter Heinrich Heine (1797-1856) zählt zu den herausragendsten und gleichzeitig kontroversesten Persönlichkeiten seiner Zeit, welche später in den deutschen literarischen Kanon ...aufgenommen wurden. Die Rezeption Heines in der Vergangenheit zeichnet sich durch wiederholte Ablehnung aus, da sowohl die Gesellschaft als auch Kritiker seinen politischen und religiösen Ansichten eher negativ gegenüberstanden. Der vorliegende Beitrag konzentriert sich auf die Rezeption Heinrich Heines im slowenischen ethnischen Gebiet bis zum Jahr 1860 und gliedert sich in drei Kapitel. Das erste Kapitel untersucht Heines Rezeption vor dem Jahr 1856, das zweite Kapitel beleuchtet seine Rezeption im Todesjahr des Dichters 1856, während sich das dritte Kapitel der Rezeption zwischen den Jahren 1857 und 1860 widmet. Die Studie umfasst eine Analyse von Veröffentlichungen in insgesamt neun journalistischen Organen, die zu dieser Zeit auf dem slowenischen ethnischen Gebiet herausgegeben wurden. Die untersuchten Zeitungen sind die Laibacher Zeitung samt ihren Beilagen, dem Intelligenzblatt zur Laibacher Zeitung, dem Illyrischen Blatt und den Blättern aus Krain, die Carinthia, die Carniolia, die Kmetijske in rokodelske novice, die Zgodnja danica sowie die literarische Zeitschrift Slovenski glasnik. Heines Präsenz ist bis zum Jahr 1860 in nahezu allen genannten Zeitungen mindestens einmal nachweisbar, mit Ausnahme der literarischen Zeitschrift Slovenski glasnik. Schlüsselwörter: Heinrich Heine, slowenisches ethnisches Gebiet, literarische Rezeption, von 1834 bis 1860, deutsche Literatur Nemski pesnik Heinrich Heine (1797-1856) je bil ze za casa svojega zivljenja ena najkontroverznejsih osebnosti, ki so kasneje postale del nemskega literarnega kanona. Zgodovinsko recepcijo Heineja je veckrat zaznamovalo zavracanje pesnika, saj druzba in kritiki niso bili naklonjeni njegovim razlicnim prepricanjem, predvsem njegovim politicnim in verskim staliscem. Pricujoci prispevek se osredotoca na recepcijo Heineja na Slovenskem do leta 1860 in je razdeljen na tri poglavja. Prvo poglavje raziskuje recepcijo Heineja na Slovenskem pred letom 1856, ko se je Heine prvic pojavil v casopisju in je njegova prisotnost v slovenskem prostoru postopoma narascala. Drugo poglavje se osredotoca na recepcijo v letu njegove smrti, 1856, pri cemer v srediscu raziskave stojita dva nekrologa. Prvi je bil objavljen v casopisu Laibacher Zeitung, medtem ko je drugi, edini izrazito negativni zapis o Heineju v tistem casu, izsel v casopisu Zgodnja danica. Tretje poglavje obravnava recepcijo med letoma 1857 in 1860. Recepcija Heineja je bila v tem casu pretezno pozitivna, z izrazitim poudarkom na pozitivnem vrednotenju Heinejeve poezije in sloga pisanja. Raziskava zajema analizo izdaj devetih publicisticnih organov, ki so v tem obdobju izhajali na Slovenskem. Pregledali smo casopise Laibacher Zeitung, njegove priloge Intelligenzblatt zur Laibacher Zeitung, Illyrisches Blatt in Blätter aus Krain, Carinthio, Carniolio, Kmetijske in rokodelske novice, Zgodnjo danico ter literarno revijo Slovenski glasnik. Heine se do leta 1860 vsaj enkrat pojavi v skoraj vseh navedenih casopisih, z izjemo literarne revije Slovenski glasnik. Kljucne besede: Heinrich Heine, slovenski prostor, literarna recepcija, od 1834 do 1860, nemska knjizevnost
In recent years, we have conducted research in various areas within the framework of the research programme Intercultural Literary Studies (No. P6-0265), especially on ethical aspects in literature, ...on the role and significance of translation as an intercultural mediation, on the literary-historical aspects of interculturality, and on the didactics of literature and translation. In the second part of this year’s double issue of Acta Neophilologica we present the latest findings of the researchers who make up the Intercultural Literary Studies programme group, and we have also invited colleagues from other programme groups as well as two colleagues from abroad.
Downstream processing of nanoplexes (viruses, virus-like particles, bacteriophages) is characterized by complexity of the starting material, number of purification methods to choose from, regulations ...that are setting the frame for the final product and analytical methods for upstream and downstream monitoring. This review gives an overview on the nanoplex downstream challenges and chromatography based analytical methods for efficient monitoring of the nanoplex production.
PDF
