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An Overview on Selection Marker Genes for Transformation of Saccharomyces cerevisiae
For genetic manipulation of yeast, numerous selection marker genes have been employed. These include prototrophic markers, markers conferring drug resistance, autoselection markers, and counterselectable markers. This chapter describes the different classes of selection markers and provides a number of examples for different applications. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - April 18, 2014 Category: Biotechnology Source Type: news

Determination of a Dynamic Feeding Strategy for Recombinant Pichia pastoris Strains
The knowledge of certain strain specific parameters of recombinant P. pastoris strains is required to be able to set up a feeding regime for fed-batch cultivations. To date, these parameters are commonly determined either by time-consuming and labor-intensive continuous cultivations or by several, consecutive fed-batch cultivations. Here, we describe a fast method based on batch experiments with methanol pulses to extract certain strain characteristic parameters, which are required to set up a dynamic feeding strategy for P. pastoris strains based on specific substrate uptake rate (q s). We further describe...
Source: Springer protocols feed by Biotechnology - April 18, 2014 Category: Biotechnology Source Type: news

Evolutionary Engineering of Yeast
Evolutionary engineering is an inverse metabolic engineering strategy which is based on increasing genetic diversity and screening large populations for desired phenotypes. This strategy is highly advantageous in certain situations over rational metabolic engineering approaches, since there is little or no requirement of detailed genetic background information for the trait of interest. Here, we describe the experimental methodology for selecting stress-resistant yeast strains via evolutionary engineering approach by either serial batch or chemostat cultivations. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - April 18, 2014 Category: Biotechnology Source Type: news

Antibody Labeling with Radioiodine and Radiometals
Antibodies have been conjugated to radionuclides for various in vitro and in vivo applications. Radiolabeled antibodies have been used in clinics and research for diagnostic applications both in vitro as reagents in bioassays and in vivo as imaging agents. Further, radiolabeled antibodies are used as direct therapeutic agents for cancer radioimmunotherapy or as tracers for studying the pharmacokinetics and biodistribution of therapeutic antibodies. Antibodies are labeled with radiohalogens or radiometals, and the choice of candidate radionuclides for a given application is dictated by their emission range and half-life. Th...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Polymer Nanoparticle-Based Controlled Pulmonary Drug Delivery
The development of novel formulations for controlled pulmonary drug delivery purposes has gained remarkable interest in medicine. Although nanomedicine represents attractive concepts for the treatment of numerous systemic diseases, scant information is available on the controlled drug release characteristics of colloidal formulations following lung administration, which might be attributed to the lack of methods to follow their absorption and distribution behavior in the pulmonary environment. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Microneedle-Iontophoresis Combinations for Enhanced Transdermal Drug Delivery
It has recently been proposed that the combination of skin barrier impairment using microneedles (MNs) coupled with iontophoresis (ITP) may broaden the range of drugs suitable for transdermal delivery as well as enabling the rate of delivery to be achieved with precise electronic control. However, few reports exist on the combination of ITP with in situ drug-loaded polymeric MN delivery systems. Our in vitro permeation studies revealed that MN enhances transdermal drug delivery. The combination of dissolving MN and ITP did not further enhance the extent of delivery of the low molecular weight drug ibuprofen sodium after sh...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Microfluidic-Based Manufacture of siRNA-Lipid Nanoparticles for Therapeutic Applications
A simple, efficient, and scalable manufacturing technique is required for developing siRNA-lipid nanoparticles (siRNA-LNP) for therapeutic applications. In this chapter we describe a novel microfluidic-based manufacturing process for the rapid manufacture of siRNA-LNP, together with protocols for characterizing the size, polydispersity, RNA encapsulation efficiency, RNA concentration, and total lipid concentration of the resultant nanoparticles. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Recombinant Stem Cells as Carriers for Cancer Gene Therapy
Recent studies have shown the ability of mesenchymal stem cells (MSCs) to migrate toward and engraft into the tumor sites, which provides a potential for their use as carriers for cancer gene therapy. Here, we describe the strategies of using MSCs as carriers for cancer gene therapy using a nonviral transfection method. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Synthesis of Thermoresponsive Polymers for Drug Delivery
A protocol for synthesizing thermosensitive copolymers of N-isopropylacrylamide (NIPAM) and N-vinylpyrrolidone (VP), cross-linked with N,N′-methylene-bis-acrylamide (MBA) has been described in this chapter. The copolymers have been formed at different concentrations of NIPAM and VP and at two different temperatures (70 °C and 30 °C). The lower critical solution temperature (LCST) of the samples has been measured, and the size of the particles formed with the highest concentration of NIPAM and lowest concentration of VP (MG1 and NG1) has been measured at three different temperatures of 25 °C, 35 °C, an...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Ultrasound-Directed, Site-Specific Gene Delivery
With the implementation of gene therapy looming in the near term, an effective delivery system using noninvasive, nonviral-mediated methods appears as an attractive option. This novel platform technology uses gas-filled, ultrasound-directed acoustic microspheres for both diagnostic imaging and therapy and yet may provide a key component for future success in the pursuit of single-gene replacement therapy. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

A Method for Screening Mitochondrial Fusogenic Envelopes for Use in Mitochondrial Drug Delivery
Various types of mitochondrial dysfunctions have been implicated in a variety of human diseases. This suggests that mitochondria would be promising therapeutic drug targets and mitochondrial therapy would be expected to be useful for the treatment of various diseases. We have already reported the development of a MITO-Porter, a liposome-based nano-carrier that delivers its cargo to mitochondria via a membrane-fusion mechanism. In our strategy for delivering cargos to mitochondria using a MITO-Porter, the carriers must fuse with the organelle membrane. Here we report on methodology for screening various types of lipid envel...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Current Status and Future Prospects of Drug Delivery Systems
This is an overview of the current drug delivery systems (DDS) starting with various routes of drug administration. Various drug formulations are then described as well as devices used for drug delivery and targeted drug delivery. There has been a considerable increase in the number of new biotechnology-based therapeutics. Most of these are proteins and peptides, and their delivery presents special challenges. Cell and gene therapies are sophisticated methods of delivery of therapeutics. Nanoparticles are considered to be important in refining drug delivery; they can be pharmaceuticals as well as diagnostics. Refinements i...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Intrathecal Delivery of Analgesics
Targeted intrathecal (IT) drug delivery systems (IDDS) are an option in algorithms for the treatment of patients with moderate to severe chronic refractory pain when more conservative options fail. This therapy is well established and supported by several publications. It has shown efficacy and is an important tool for the treatment of spasticity, and both cancer and nonmalignant pain. Recent technological advances, new therapeutic applications, reported complications, and the costs as well as maintenance required for this therapy require the need to stay up-to-date about new recommendations that may improve outcomes. This...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Intranasal Delivery of Chitosan–siRNA Nanoparticle Formulation to the Brain
Neurodegeneration is characterized by a progressive loss of neuron structure and function. Most neurodegenerative diseases progress slowly over the time. There is currently no cure available for any neurodegenerative disease, and the existing therapeutic interventions only alleviate the symptoms of the disease. The advances in the drug discovery research have come to a halt with a lack of effective means to deliver drugs at the targeted site. In addition, the route of delivering the drugs is equally important as most invasive techniques lead to postoperative complications. This chapter focuses on a non-invasive, intranasal...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news

Synthesis of a Smart Nanovehicle for Targeting Liver
A protocol for the synthesis of a smart drug delivery system based on gold nanoparticles has been described in this chapter. The synthesized drug delivery system has been shown to release the bioactive material in response to an intracellular stimulus (glutathione concentration gradient) and hence shown to behave in an intelligent manner. Gold nanoparticles have been employed as the core material with the surface functionalities of thiolated PEG. PEG owing to its non-immunogenicity and non-antigenicity would impart considerable stability and longer in vivo circulation time to the gold nanoparticles. The end groups of PEG c...
Source: Springer protocols feed by Biotechnology - February 25, 2014 Category: Biotechnology Source Type: news