Proizvodnja cjepiva protiv gripe - dosezi i izazovi Slivac, Igor; Buljubašić, Ena; Gaurina Srček, Višnja ...
Hrvatski časopis za prehrambenu tehnologiju, biotehnologiju i nutricionizam,
01/2021, Volume:
15, Issue:
3-4
Journal Article
Open access
Gripa (ili influenca) je virusna, infektivna i respiratorna bolest koja godišnje zarazi do milijardu ljudi diljem svijeta. Pojavljuje se svake godine kao sezonska gripa, no u povijesti se svakih ...nekoliko desetaka godina javljala i kao pandemija. Njezin nepovoljan učinak na pojedinca i
društvo može se spriječiti upotrebom cjepiva, tj. cijepljenjem. Prvo odobreno cjepivo protiv gripe proizvedeno je u embrioniranom kokošjem jajetu prije skoro 80 godina, a po sličnom tehnološkom načelu proizvode se i današnja cjepiva. Napredak u razumijevanju stanične biologije te unaprjeđenje proizvodnih postupaka pomoću kulture stanica omogućilo je stručnjacima prijelaz s konvencionalnih na nove, produktivnije i brže proizvodne postupke. U radu se najprije razmatra epidemiloški aspekt gripe, struktura virusa gripe, a zatim dosezi i izazovi u suvremenoj proizvodnji cjepiva protiv gripe. U završnom dijelu ističu se potrebe i trendovi u izradi univerzalnog cjepiva protiv gripe.
Influenza (flu) is a viral, contagious, respiratory disease that annually infects up to a billion people worldwide. It appears each year as a seasonal flu, but through history, it appeared as pandemic illness once in every several decades. Its harmful effects on human health and consequently to the society can be prevented by vaccination. The first approved influenza vaccine was made almost 80 years ago using embryonated hen eggs and the same technology is still in use for most available vaccines today. The development of cell biology and improvement of cell culture manufacturing techniques enabled the transition from conventional to novel, more efficient vaccine production methods. In this work we describe the epidemiology of influenza, influenza virus structure as well as current influenza vaccine manufacturing approaches. A brief overview of universal vaccine development and its urgency is discussed in the final section of the paper.
U procesima s kulturama životinjskih stanica koristi se medij za uzgoj koji sadrži osnovne nutrijente poput glukoze, amino-kiselina, soli te hormone i faktore rasta iz seruma životinjskog porijekla. ...Zbog nedostataka primjene seruma u procesima s kulturama stanica (npr. rizik prijenosa virusa, varijacije u sastavu, otežana izolacija proizvoda) pristupilo se razvoju medija bez seruma u koje se često dodaju i hidrolizati proteina biljnog porijekla. Hidrolizati sadrže aminokiseline, oligopeptide, lipide, vitamine i ostale spojeve koji potiču proliferaciju i produktivnost stanica u kulturi. U ovom radu dan je kratak pregled priprave i najvažnijih bioloških svojstava proteinskih hidrolizata s naglaskom na primjenu biljnih hidrolizata kao sastojaka medija za uzgoj životinjskih stanica u proizvodnji biofarmaceutika.
Culture media containing essential nutrients like glucose, amino-acids and salts, as well as hormones, growth factors added by animal blood serum is required for bioprocesses with animal cells. Due to the disadvantages of serum use (e.g. risk of virus transmission, variation in composition, complex product isolation), the development of serum-free media supplemented with plant protein hydrolysates took place about two decades ago.The hydrolysates contain amino acids, oligopeptides, lipids, vitamins and other compounds that promote cell culture proliferation and productivity. This paper gives a brief overview of the preparation and the most important biological properties of protein hydrolysates with emphasis on the use of plant hydrolysates as components of animal cell culture media in the production of biopharmaceuticals.
This is the first report about flaxseed protein hydrolysates applied as media supplements in CHO cell culture. The hydrolysates were produced by three separate enzymatic digestions of proteins ...isolated from flaxseed oil cake. The enzymes used were Alcalase, Neutrase, and Protamex, and the most efficient hydrolysis was achieved with Alcalase. The three hydrolysates were first tested as a partial substitute for serum in basal media in order to evaluate their effects on the adherent IgG-producing CHO cell line. The cells that grew in such media reached higher density than the cells in media supplemented with serum only. Consequently, the increased cell number improved the final IgG titer. In the next experiment, the impact of hydrolysates was evaluated in suspension CHO culture adapted to chemically defined media. In this preliminary investigation, the cells showed no response to the hydrolysate addition concerning the growth rate and productivity. Despite this outcome, we speculate that low molecular mass components in the hydrolysates, besides nutritive, may have a cell-protective function.
Flaxseed meal, a byproduct of flaxseed oil extraction, was treated as low-value agrowaste for a long time despite its high protein content. Flaxseed meal has recently garnered increasing interest as ...a source of proteins and other bioactive compounds with positive impacts on human health. The aim of this study was to investigate the
in vitro
biological potential of flaxseed protein hydrolysates (FPH). Three FPHs were prepared using three hydrolytic enzymes: Alcalase, Neutrase and Protamex. The molecular weight profile of peptides contained in the hydrolysates was determined by size exclusion chromatography (SEC). The oxygen radical absorbance capacity (ORAC) assay was used to determine the peptide antioxidant capacity, while proliferative effects were studied in two cell lines: HeLa and HaCaT. The latter was also used to determine the protective effect of the FPH during induced oxidative stress. Alcalase showed the highest proteolytic activity, while the produced flaxseed protein hydrolysate (FPH-A) exhibited the strongest antioxidant potential. FPH-A had cytotoxic effects at 10 mg/mL in HeLa cells, but it stimulated HaCaT cell growth. Moreover, a mild protective effect of FPH-A was detected in HaCaT cells after induction of oxidative stress. Flaxseed protein hydrolysates obtained by Neutrase (FPH-N) and Protamex (FPH-P) have less pronounced or no potential at all, with respect to their antioxidative or antiproliferative activity. Therefore, to increase value-added utilization of flaxseed meal we suggest further research on hydrolysate obtained by Alcalase.
Food proteins from different sources can provide beneficial effects on human health by releasing the bioactive peptides that are integral part of their native structure. In this study, we tested the ...biological potential of hempseed protein hydrolysates (HPHs) obtained from hempseed cake protein isolate. The HPHs were prepared by enzyme hydrolysis using three different proteases of microbial origin: Alcalase®, Neutrase® and Protamex®. The antioxidant activity of the obtained hydrolysates was determined by oxygen radical absorbance capacity (ORAC) assay, while the proliferative effects on normal (HaCaT) and cancer (HeLa) cells were determined by the CellTiter 96
®
AQ
ueous
One Solution Reagent (MTS) assay. HPHs showed dose-dependent antiproliferative effects on HeLa cells and stimulatory effects on the proliferation of HaCaT cells. HPH obtained by Neutrase
®
(HPH-N) showed the highest antioxidant activity expressed as an ORAC value. The protective effect of HPH-N on H
2
O
2
-induced oxidative stress in normal and cancer cells was evaluated and 1 mg/mL of HPH-N significantly reduced the formation of intracellular reactive oxygen species (ROS) in both cell lines. The obtained results indicate the benefits of HPHs as potential natural antioxidants for the food industry and contribute to the growing trend of utilizing hempseed by-products.
This paper presents the levels of sulphur, polycyclic aromatic hydrocarbons (PAHs), and potentially toxic trace elements in soils surrounding the Plomin coal-fired power plant (Croatia). It used ...domestic superhigh-organic-sulphur Raša coal from 1970 until 2000. Raša coal was characterised by exceptionally high values of S, up to 14%, making the downwind southwest (SW) area surrounding the power plant a significant hotspot. The analytical results show that the SW soil locations are severely polluted with S (up to 4%), and PAHs (up to 13,535ng/g), while moderately with Se (up to 6.8mg/kg), and Cd (up to 4.7mg/kg). The composition and distribution pattern of PAHs in the polluted soils indicate that their main source could be airborne unburnt coal particles. The atmospheric dispersion processes of SO2 and ash particles have influenced the composition and distribution patterns of sulphur and potentially toxic trace elements in studied soils, respectively. A possible adverse impact of analysed soil on the local karstic environment was evaluated by cytotoxic and genotoxic methods. The cytotoxicity effects of soil and ash water extracts on the channel catfish ovary (CCO) cell line were found to be statistically significant in the case of the most polluted soil and ash samples. However, the primary DNA-damaging potential of the most polluted soil samples on the CCO cells was found to be within acceptable boundaries.
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•Soil from coal plant is highly polluted with S (max 4%) and PAHs (max 13,535ng/g).•Selenium and Cd are significantly enriched in the most polluted soil.•The most polluted soil and bottom ash induced significant cytotoxicity on CCO cells.•The most polluted soil induced no significant genotoxicity on CCO cells.
Nastojanje suvremene prehrambene industrije je što učinkovitije iskoristiti organske ostatke nastale preradom sirovina u prodajne proizvode. Primjer takvog organskog ostatka je i uljna pogača, koja ...zaostaje nakon ekstrakcije ulja iz sjemenki lana, a zbog svog sastava proteina predstavlja potencijalnu sirovinu za primjenu u tehnologiji uzgoja stanica. Stoga je iz uljne pogače lana izoliran proteinski izolat
te je ispitan njegov utjecaj na rast CHO-E i HEK-293T staničnih linija,
kao i na produktivnost CHO-E stanične linije. Dokazano je da dodatak proteinskog izolata u hranjivi medij u vrlo niskim koncentracijama (0,1 g
L-1 i/ili 0,2 g L-1) ima pozitivan učinak na rast obje stanične linije,
kao i na produktivnost CHO-E stanične linije. Međutim, dodatak većih koncentracija izolata od navedenih ne utječe pozitivno na rast obje stanične linije, odnosno na njih djeluje inhibitorno.
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