Springer protocols feed by Biotechnology 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Bacterial Systems for Gene Delivery to Systemic Tumors
Certain bacteria have emerged as biological gene vectors with natural tumor specificity, capable of specifically delivering genes or gene products to the tumor environment when intravenously (i.v.) administered to rodent models. Here, we describe procedures for studying this phenomenon in vitro and in vivo for both invasive and noninvasive bacteria suitable for exploitation as tumor-specific therapeutic delivery vehicles, due to their ability to replicate specifically within tumors and/or mediate bacterial-mediated transfer of plasmid DNA to mammalian cells (bactofection). (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news
A Method for Evaluating Nanoparticle Transport Through the Blood–Brain Barrier In Vitro
Blood–brain barrier (BBB) represents a formidable barrier for many therapeutic drugs to enter the brain tissue. The development of new strategies for enhancing drug delivery to the brain is of great importance in diagnostics and therapeutics of central nervous system (CNS) diseases. In this context, nanoparticles are an emerging class of drug delivery systems that can be easily tailored to deliver drugs to various compartments of the body, including the brain. To identify, characterize, and validate novel nanoparticles applicable to brain delivery, in vitro BBB model systems have been developed. In this work, we desc...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news
Applications of Chitosan Nanoparticles in Drug Delivery
We have reviewed the binding affinities of several antitumor drugs doxorubicin (Dox), N-(trifluoroacetyl) doxorubicin (FDox), tamoxifen (Tam), 4-hydroxytamoxifen (4-Hydroxytam), and endoxifen (Endox) with chitosan nanoparticles of different sizes (chitosan-15, chitosan-100, and chitosan-200 KD) in order to evaluate the efficacy of chitosan nanocarriers in drug delivery systems. Spectroscopic and molecular modeling studies showed the binding sites and the stability of drug–polymer complexes. Drug–chitosan complexation occurred via hydrophobic and hydrophilic contacts as well as H-bonding network. Chitosan-100 KD...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news
Self-Assembling Peptide-Based Delivery of Therapeutics for Myocardial Infarction
Drug and cell delivery systems could be modulated to serve as instructive microenvironments in regenerative medicine. Towards this end, several synthetic biomaterials have been developed to mimic the natural extracellular matrix (ECM) for therapeutic use. These include synthetic polymers, decellularized ECM, self-assembling polymers, and cell-responsive hydrogels with varied applications. Here, we describe the development of a self-assembling peptide hydrogel and its potential use as a cell and growth factor delivery vehicle to the infarcted heart in a rodent model of myocardial infarction. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news
Designing Media for Animal Cell Culture: CHO Cells, the Industrial Standard
The success of culturing CHO cells solely depends on functionality of the used media. Cell culture technology is more than 50 years old, and the knowledge of cell requirements increased steadily. In the beginning, animal-sourced components were the key to growth. Nowadays state-of-the-art media do not contain any animal or naturally sourced components. The compositions are based on scientific awareness of the needs of the cells. The result is high lot-to-lot consistency and high performance. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
Tools for High-Throughput Process and Medium Optimization
Shaking technology in combination with small-scale disposable plastic vessels has become a notable bioprocess optimization tool widely exploited for cells grown in suspension. This chapter focuses on the two most accommodating culture systems: 50 mL centrifugation tubes and 96-deepwell plates. Used by many laboratories for routine passaging of suspension cultures and all types of optimization experiments as flexible culture system, the 50 mL shake tubes are the preferred vessels for manual manipulations, while microtiter plates are the ideal containers when automatic liquid handling systems are available. Both culture syst...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
Microbioreactors and Scale-Down Models: Growth of CHO Cells Using the Pall Micro24 MicroReactor System
Microbioreactors are increasingly used within animal cell biotechnology to grow mammalian cells for cell line screening and to facilitate process development. Many such devices have been reported in the literature, but only a small number are available commercially. Microbioreactors range in complexity from simple plate-based systems to complex automated parallel bioreactors designed to enable the meaningful scale-down of conventional bioprocesses. The Micro24 MicroReactor system (Pall Life Sciences) fits between these extremes providing 24× 7 mL parallel “bioreactors” with individual monitoring and contr...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
Evaluation of Solid and Porous Microcarriers for Cell Growth and Production of Recombinant Proteins
Microcarrier technology opened new perspectives for anchorage-dependent cell culture, by providing increased surface areas for cell adhesion and proliferation, and therefore improving both cell and product yields obtained in these cultures. The establishment of a successful microcarrier culture depends on many factors, such as the type of microcarrier, the cells, and the culture conditions. In this chapter, the basic steps required for the evaluation and optimization of a microcarrier culture for the purpose of production of recombinant proteins are described, for both solid and porous microcarriers. (Source: Springer prot...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
DNA Profiling and Characterization of Animal Cell Lines
The history of the culture of animal cell lines is littered with published and much unpublished experience with cell lines that have become switched, mislabelled, or cross-contaminated during laboratory handling. To deliver valid and good quality research and to avoid waste of time and resources on such rogue lines, it is vital to perform some kind of qualification for the provenance of cell lines used in research and particularly in the development of biomedical products. DNA profiling provides a valuable tool to compare different sources of the same cells and, where original material or tissue is available, to confirm th...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
Scalable Transient Protein Expression
Transient transfection is a well-established method to rapidly express recombinant proteins from mammalian cells. Accelerating activity in biotherapeutic drug development, demand for protein-based reagents, vaccine research, and large initiatives in structural and functional studies of proteins have propelled the need to generate moderate to high amounts of recombinant proteins and other macromolecules in a flexible and rapid manner. Progress over the last 10–15 years has demonstrated that transient transfections can be reliably and readily scaled up to handle milliliters to tens of liters of cells in suspension cult...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
Scalable Transient Gene Expression in Adherent Mammalian Cells Using Polyethylenimine
Transient gene expression (TGE) has become a powerful tool over the last decade, as time to market is a key factor in biopharmaceutical industry. TGE is used to produce small amounts of recombinant protein for functional and structural studies. To broaden the applicability of the method, the scale-up of TGE has been in scope of the scientific community over the last decade. Culture volumes of up to 100 l have been transfected, but there are still bottlenecks regarding the compatibility of the transfection method and media composition as well as media exchange or dilution required after transfection. Cell immobilization on ...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
Insect Cell Line Development Using Flp-Mediated Cassette Exchange Technology
Traditional cell line development is quite laborious and time-consuming as it is based on the random integration of the gene of interest which leads to unpredictable expression behavior. In opposition, recombinase-mediated cassette exchange systems represent a powerful genetic engineering approach, allowing site-specific insertion of recombinant genes into pre-tagged genomic loci with superior expression characteristics, thus bypassing the need for extensive clone screening and shortening the development timelines. Such systems have not been widely implemented in insect cell lines used for the production of recombinant pro...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
High-Throughput Synchronization of Mammalian Cell Cultures by Spiral Microfluidics
The development of mammalian cell cycle synchronization techniques has greatly advanced our understanding of many cellular regulatory events and mechanisms specific to different phases of the cell cycle. In this chapter, we describe a high-throughput microfluidic-based approach for cell cycle synchronization. By exploiting the relationship between cell size and its phase in the cell cycle, large numbers of synchronized cells can be obtained by size fractionation in a spiral microfluidic channel. Protocols for the synchronization of primary cells such as mesenchymal stem cells, and immortal cell lines such as Chinese hamste...
Source: Springer protocols feed by Biotechnology - December 4, 2013 Category: Biotechnology Source Type: news
Molecular Targeted Viral Nanoparticles as Tools for Imaging Cancer
Viral nanoparticles (VNPs) are a novel class of bionanomaterials that harness the natural biocompatibility of viruses for the development of therapeutics, vaccines, and imaging tools. The plant virus, cowpea mosaic virus (CPMV), has been successfully engineered to create novel cancer-targeted imaging agents by incorporating fluorescent dyes, polyethylene glycol (PEG) polymers, and targeting moieties. Using straightforward conjugation strategies, VNPs with high selectivity for cancer-specific molecular targets can be synthesized for in vivo imaging of tumors. Here we describe the synthesis and purification of CPMV-based VNP...
Source: Springer protocols feed by Biotechnology - November 20, 2013 Category: Biotechnology Source Type: news
Phage-Displayed Single Domain Antibodies as Recognition Elements
The variable domains of antibodies can be expressed as a fusion with pIII, a minor coat protein of the bateriophage M13, for the generation of phage-display antibody reagents. The phage-display system is routinely used to enrich for recombinant antibodies against a specific target antigen from highly diverse naïve and immune libraries. Often once binders are selected, they are expressed as soluble proteins; however, it can be advantageous to use the phage-displayed antibody fragment as a reagent in binding assays. The repeating subunits of the viral capsid allows for significant signal amplification of binding events ...
Source: Springer protocols feed by Biotechnology - November 20, 2013 Category: Biotechnology Source Type: news
