Recovery of virus producing NDV vaccine by Tangential Flow Filtration (TFF)

Currently, virus separation process is mainly conducted by using ultracentrifugation or sucrose gradient electrophoresis methods. Both methods however have several drawbacks wherein the ultracentrifugation method requires very high rotational speed to separate the virus while the sucrose gradient electrophoresis method is very time consuming. Alternative to both methods are by using Tangential Flow Filtration (TFF). In this study, we have separated and concentrated the Newcastle Disease Virus (NDV) harvested from embryonic specific pathogen free (SPF) eggs using TFF method by utilizing microfilter membrane with pore size of 0.45μm. Hollow fiber membrane was selected as filter because of their chemical and thermal stability. The study was conducted according to the design developed using Taguchi method which consists of selected three parameters and two levels of factors. The results revealed that higher pressure input (Pin), lower pressure output (Pout), and higher virus concentration led to higher virus titer. Optimum Trans-Membrane-Pressure (TMP) value of 15 psi and virus concentration of 28% had given the maximum titer of the virus which was 512 haemagglutination assay (HA) unit

Review on transfersomes : Promising carrier for transdermal drug delivery

The transdermal drug delivery systems (TDDS) facilitate over the traditional techniques by including the ability to deliver a drug more selectively to a specific point; more straightforward, more accurate, and less frequent dosing besides decreasing variability in systemic drug concentrations; absorption that is more consistent with the site and mechanism of action; and minimized toxic metabolites. Moreover, these are painless and self-administered delivery systems that can increase patient compliance and provide a controlled release of the medications. However, the greatest challenge of TDDS is the barrier function of stratum corneum (SC), the outermost layer of the skin which allows only unionised molecule and molecule with molecular weight less than 500 Da to pass through it. As a result, this method can only be used to provide a restricted number of medications. Now a days, some of the possible alternatives to this challenge are chemical permeation enhancher, iontophoresis, sonophoresis, microneedles, electrophoresis and vesicular system such as liposome, transfersome, ethosome etc. Transferosomes appear to be one of the most promising of these techniques, as they feature a bilayered shape that allows lipophilic and hydrophilic drugs, as well as amphiphilic drugs, to be encapsulated with better penetration efficiencies than typical liposomes. Transfersomes are elastic in nature, which can deform and squeeze themselves as an undamaged vesicle through narrow pores that are significantly smaller than its size. This review discusses the concept of transfersomes, composition of transfersomes, mechanism of action and various preparation techniques. Also, this review focuses on recent applications of transfersomes along with their benefits and drawbacks. Additionally, this review sheds light on future research scope to motivate the researchers for future outstanding research works

Optimization of process conditions for high cell density proliferation of DF-1 cells in bioreactor

The purpose of this study was to optimize agitation speed, microcarrier concentration and pO2 levels in bioreactor for culturing DF-1cells in DMEM media supplemented with 7% fetal bovine serum. Statistical analysis has shown that the model derived from the results was proven to be significant and pO2 level turned out as the most significant parameter in the experiment. As predicted by analysis, cells grew up to 1.190 x 106 cells/ml when moderate agitation speed, microcarrier concentration, and pO2 level are applied

Comparison of growth rate and viability of DF-1 cell line in different culture media

One of the commercial cell line that gained attention for the study of viral vaccine was an immortalized cell line derived from chicken embryo fibroblast (CEF) cells. Unlike CEF cells, DF-1 cells have higher proliferation rate and also can be subcultured infinite number of times. In this study, the growth profile of DF-1 cell line was investigated by undergoing series of optimization process by testing it out on several culture media and also by manipulating the components of culture media which were sodium bicarbonate and serum. At the end of this study it has been figured out that DF-1 achieved the highest cell culture density in Dulbecco’s Modification of Eagle’s Medium( DMEM) and with 1.24 g/L of NaHCO3 and 5.5 % serum, it yielded the shortest doubling time

Growth study of DF-1 cell line in microcarrier bioreactor

Animal cell culture is fundamental in the manufacturing of many biological products. Biological products produced in animal cell cultures include vaccines, enzymes, hormones, antibodies and anticancer agents. However majority of animal cells is anchorage-dependent and requires attachment to a surface for their growth. Microcarrier is a small bead used to provide surface for the culture of anchorage-dependent animal cells. Microcarrier culture has very large surface area to volume ratio, thus able to produce larger number of cells when compared to conventional monolayer cultures which uses multiple glass or plastic bottles. This research was carried out mainly to study the growth performance of DF-1 cell culture by using microcarriers in a 1-L litre stirred tank bioreactor and compares it with conventional monolayer cultures grown in T-flasks bioreactor. At the end of the study it has been found that the maximum viable cell concentration achieved by the microcarrier culture was 1.315 × 106 cells/ml which was higher than maximum viable cell concentration achieved by the cell culture in T-flasks which was 1.25 × 106 cells/ml

Surface modification of polycaprolactone (PCL) microcarrier for performance improvement of human skin fibroblast cell culture

Polycaprolactone (PCL) has many advantages for use in biomedical engineering field. In the present work PCL microcarriers of 150-200 µm were fabricated using oil-in-water (o/w) emulsification coupled with solvent evaporation method. The surface charge of PCL microcarrier was then been improved by using ultraviolet/ozone treatment to introduce oxygen functional group. Immobilisation of gelatin onto PCL microspheres using zero-length crosslinker provides a stable protein-support complex, with no diffusional barrier which is ideal for mass processing. The optimum concentration of carboxyl group (COOH) absorbed on the surface was 1495.9 nmol/g and the amount of gelatin immobilized was 1797.3 µg/g on UV/O3 treated microcarriers as compared to the untreated (320 µg/g) microcarriers. The absorption of functional oxygen groups on the surface and the immobilized gelatin was confirmed with the attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR) and the enhancement of hydrophilicity of the surface was confirmed using water contact angle measurement which decreased (86.93o – 49.34o) after UV/O3 treatment and subsequently after immobilisation of gelatin. The attachment and growth kinetics for human skin fibroblast cell (HSFC) showed that adhesion occurred much more rapidly for gelatin immobilised surface as compared to untreated PCL and UV/O3 PCL microcarrier