By Anthony P. Green (auth.), Douglas B. Lowrie MD, Robert G. Whalen MD (eds.)
In DNA Vaccines: tools and Protocols, state of the art evaluate articles by means of top specialists summarize the best way to increase and hire the hugely promising new DNA vaccines, what medical effects may be anticipated from their use, and what's recognized approximately how they paintings. Key themes variety from vaccine layout and building to training and supply equipment, together with using classical adjuvants, "genetic adjuvants," and the immunostimulatory homes of DNA and chosen oligonucleotide sequences. a number of individuals offer strategic rules on antigen engineering and describe the relatively novel functions of DNA vaccine method that experience lately emerged. additionally, there's a dialogue of dendritic cells and antigen-presenting cells, the knowledge of whose activities holds nice promise for modulating the immune reaction, and therefore for treating disorder. state of the art and complete, DNA Vaccines: tools and Protocols offers a wide landscape of the tools and pondering from which the vaccines of day after today will evolve, and so constitutes a useful sourcebook for either specialists constructing new purposes and beginners who are looking to achieve mastery of the innovations and difficulties concerned.
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Additional info for DNA Vaccines: Methods and Protocols
Sample text
In case of large numbers of clones, use an automated device for the isolation of DNA in a 96-well format (BioRobot 9600, Qiagen). 4. Identify appropriate cell clones by comparing them and selecting those with high plasmid yield and a proper plasmid isoform distribution for further production steps (Fig. 2). Production of Plasmid DNA 17 Fig. 2: Comparison of plasmid pUC21 DNA produced in different E. coli K12 host cells. The upper panel shows the undigested DNA with its different isoforms. The lower panel shows as a control the same amount of DNA after an EcoRI digestion.
5 with NaOH, and sterilize by autoclaving. Dispense the medium by half-filling sterile conical flasks and add the selective antibiotic appropriate to the plasmid of interest. 3. LB agar plates: these are prepared as above but containing 15 g/L Bacto-agar and dispensed into sterile Petri dishes after addition of the antibiotic. 3. Methods 1. Streak out the transfected bacteria on two LB agar plates, one containing the recommended concentration of antibiotic and one containing double the concentration.
0. Human IL-2 expression in the supernatant of UM449 cells after transfection was measured by ELISA. Formulation A is: 12% trehalose, 2% PVP. Formulation B is: 15% trehalose, 2% PVP, 2% PEG. supernatants 48 h post-transfection. There is no significant difference in IL-2 expression between lyophilized VCL1102 and non-lyophilized control DNA. 6. In Vivo Bioassays of the Reconstituted Lyophilized DNA Formulation A panel of lyophilized VR1223 plasmid DNA formulations were evaluated for luciferase expression in mice.