The Philips Lecture, 1981 - Targets in biotechnology

Established biotechnologies overcome cost handicaps, particularly aqueous process dilution, through unique advantages. Ancient fermentations confer psychopharmacological, palatable or nutritional qualities on drinks and foods. Modern biotechnologies depend on bacterial versatility (sewage disposal), enzyme specificity (hydrolases), high value of complex products (antibiotics, vaccines), chirality (amino acids), high yield (citric acid), or, rarely, process intensity (glucose isomerase). Advances in recombinant DNA have already given valuable human proteins. Numerous new targets include hormones, blood proteins and antibodies. Production techniques will include cell fusion and animal cell culture, and new hosts, some eukaryotic, for human genes. Enhanced production of cells, organelles and enzymes will follow, partly to take advantage of the industrial potential of immobilization. Later still, improved engineering and new biological processes should allow competitive production of fine chemicals, protein and other foods, bulk chemicals and fuels. Some products will require simultaneous change and advance in agriculture: interspecific genetic manipulation, plant cell culture and other modern developments will facilitate these.

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A Brief Concept of Cell Culture: Challenges, Prospects and Applications

10.5772/intechopen.99387 ◽

2021 ◽

Author(s):

Md. Salauddin

Keyword(s):

Cell Culture ◽

Recombinant Dna ◽

Vaccine Development ◽

Culture Media ◽

Animal Cell ◽

Laboratory Research ◽

Cell Infection ◽

Ph Indicator ◽

Whole Process

Cell culture is an in vitro technique in which cells, tissues, or organs (animal origin) are artificially grown with the support of an artificial environment that encompasses culture medium, CO2 level, pH indicator, temperature keeping tissues alive and growing appropriately. Organ culture, Primary explant culture, and Cell culture among them cell culture widely used for the understanding of cell growth, normal functions, identification of growth factors, viral vaccine development, recombinant DNA (rDNA) technology, and immunobiological research. Due to high feasibility, cell culture practices highly demandable in the pharmaceutical industry. As well as animal cell culture used in laboratory research to study the cytotoxicity of new drug metabolic studies, aging, therapeutic proteins, the effects of drugs and toxic compounds on the cells and mutagenesis and carcinogenesis. There are a lot of issues in cell culture, Mycoplasma is one of the major. During cell culture, a single antibiotic often cannot kill the mycoplasma. Besides, culture media, pH indicator, incubation, cryopreservation, thawing, passaging of cells, and trypsinization have a great impact on cell culture. This chapter will help the reader to understand the whole process of cell culture and its applications, which will take them one step forward in their virology and cell culture research along with inspiration. This chapter also aids in the concept of cell count, cell suspension, CCF measurement, MOI (Multiplicity of Infection), and cell infection. Eventually, the reader will get a crystal clear concept of cell culture.

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A Review: Klebsiella pneumoniae, Klebisellaoxytoca and Biotechnology

ROMANIAN BIOTECHNOLOGICAL LETTERS ◽

10.25083/rbl/26.3/2567.2586 ◽

2021 ◽

Vol 26 (3) ◽

pp. 2567-2586

Author(s):

CATERINA TOMULESCU ◽

◽

MIȘU MOSCOVICI ◽

IRINA LUPESCU ◽

ROXANA MĂDĂLINA STOICA ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Genetic Manipulation ◽

Klebsiella Oxytoca ◽

Cost Effective ◽

Biochemical Properties ◽

High Yield ◽

Natural Environments ◽

Klebsiella Species ◽

Bulk Chemicals ◽

Plant Growth Promoting Activities

Biotechnology, molecular biology and genetic engineering, and bioprospecting play a crucial role in our common future, enabling industrially important microorganisms to ensure sustainable products (fuels, chemicals, pharmaceuticals, food, drug delivery systems, medical devices etc.) and new bioeconomic opportunities. Biotechnological applications are able to provide cost-effective green alternatives to conventional industrial processes, which are currently affecting the nature and biodiversity. Klebsiella species are among the well-studied microbes both in medicine field, as ones of the most resilient opportunistic pathogens, and in industry, due to their promising biochemical properties, and their potential as better hosts than other microorganisms, for i.e. in genetic manipulation. Klebsiella oxytoca and Klebsiella pneumoniae are ubiquitously found in natural environments, but also as commensals in the human gut, and associated with a high-resistance to the first-line antibiotics. However, these specific strains are continuously isolated and studied for different industrial purposes (i.e. bulk chemicals and biofuels production, medical diagnosis, nanoparticles and exopolysaccharides synthesis, plant growth promoting activities, bioremediation and biodegradation agents etc.), and scientific results regarding their biotechnological potential could generate big impact for global bioeconomy development. Recently, research in synthetic biology gained a lot of attention, and new techniques highlight ways to reprogramme these microbial cells in view of high-yield or high-quality new chemicals obtainment. Therefore, some scientific research niches are emerging in biotechnology, and unknown metabolic pathways and genes are identified and further studied, to provide alternative solutions to the global challenges.

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Book Review: Animal Cell Culture: A Practical Approach

Alternatives to Laboratory Animals ◽

10.1177/026119298501300415 ◽

1985 ◽

Vol 13 (4) ◽

pp. 304-304

Author(s):

Michael Balls

Keyword(s):

Cell Culture ◽

Animal Cell ◽

Animal Cell Culture ◽

Practical Approach

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Site-specific N-glycosylation analysis of animal cell culture-derived Zika virus proteins

Scientific Reports ◽

10.1038/s41598-021-84682-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Alexander Pralow ◽

Alexander Nikolay ◽

Arnaud Leon ◽

Yvonne Genzel ◽

Erdmann Rapp ◽

...

Keyword(s):

Zika Virus ◽

Animal Cell ◽

Gradient Centrifugation ◽

Viral Envelope ◽

Protein Glycosylation ◽

High Yield ◽

Structural Protein ◽

Site Specific ◽

E Protein ◽

The Impact

AbstractHere, we present for the first time, a site-specific N-glycosylation analysis of proteins from a Brazilian Zika virus (ZIKV) strain. The virus was propagated with high yield in an embryo-derived stem cell line (EB66, Valneva SE), and concentrated by g-force step-gradient centrifugation. Subsequently, the sample was proteolytically digested with different enzymes, measured via a LC–MS/MS-based workflow, and analyzed in a semi-automated way using the in-house developed glyXtoolMS software. The viral non-structural protein 1 (NS1) was glycosylated exclusively with high-mannose structures on both potential N-glycosylation sites. In case of the viral envelope (E) protein, no specific N-glycans could be identified with this method. Nevertheless, N-glycosylation could be proved by enzymatic de-N-glycosylation with PNGase F, resulting in a strong MS-signal of the former glycopeptide with deamidated asparagine at the potential N-glycosylation site N444. This confirmed that this site of the ZIKV E protein is highly N-glycosylated but with very high micro-heterogeneity. Our study clearly demonstrates the progress made towards site-specific N-glycosylation analysis of viral proteins, i.e. for Brazilian ZIKV. It allows to better characterize viral isolates, and to monitor glycosylation of major antigens. The method established can be applied for detailed studies regarding the impact of protein glycosylation on antigenicity and human pathogenicity of many viruses including influenza virus, HIV and corona virus.

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Defined MSC exosome with high yield and purity to improve regenerative activity

Journal of Tissue Engineering ◽

10.1177/20417314211008626 ◽

2021 ◽

Vol 12 ◽

pp. 204173142110086

Author(s):

Jun Yong Kim ◽

Won-Kyu Rhim ◽

Yong-In Yoo ◽

Da-Seul Kim ◽

Kyoung-Won Ko ◽

...

Keyword(s):

Cell Culture ◽

Culture Medium ◽

Density Lipoprotein ◽

High Yield ◽

Human Umbilical Cord ◽

Cell Culture Medium ◽

Human Coronary Artery ◽

Isolation Methods ◽

Regenerative Activity ◽

Yield And Purity

Exosomes derived from mesenchymal stem cells (MSCs) have been studied as vital components of regenerative medicine. Typically, various isolation methods of exosomes from cell culture medium have been developed to increase the isolation yield of exosomes. Moreover, the exosome-depletion process of serum has been considered to result in clinically active and highly purified exosomes from the cell culture medium. Our aim was to compare isolation methods, ultracentrifuge (UC)-based conventional method, and tangential flow filtration (TFF) system-based method for separation with high yield, and the bioactivity of the exosome according to the purity of MSC-derived exosome was determined by the ratio of Fetal bovine serum (FBS)-derived exosome to MSC-derived exosome depending on exosome depletion processes of FBS. The TFF-based isolation yield of exosome derived from human umbilical cord MSC (UCMSC) increased two orders (92.5 times) compared to UC-based isolation method. Moreover, by optimizing the process of depleting FBS-derived exosome, the purity of UCMSC-derived exosome, evaluated using the expression level of MSC exosome surface marker (CD73), was about 15.6 times enhanced and the concentration of low-density lipoprotein-cholesterol (LDL-c), known as impurities resulting from FBS, proved to be negligibly detected. The wound healing and angiogenic effects of highly purified UCMSC-derived exosomes were improved about 23.1% and 71.4%, respectively, with human coronary artery endothelial cells (HCAEC). It suggests that the defined MSC exosome with high yield and purity could increase regenerative activity.

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