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Adeno-associated virus-like particle (AAVLP) Heat Shock Protein (HSP) vaccine for the treatment of Vitiligo and other autoimmune diseases
Denmark (IMCC) - Aalborg University, Aalborg
Health Science and Technology, Metabolism Modifying Medicine (MMM) group
John Nieland and Jeanette Prangsgaard
English, German, Danish, Dutch
Type of Research Project
- Basic science
What is the background of the project?
In the MMM group we are working on a virus like particle vaccine (AAVLP) encompassing a part of viral protein sequences which are involved in the transfer of a virus over a cellular membrane. The AAVLP Viral peptide particle will, after immunization, induce a viral peptide specific antibody response that will block the virus from entering host cells. Therewith, the vaccine will block the infection with the virus.
What is the aim of the project?
The aim of the project is the optimization of production and purification of AAVLP viral peptide vaccine.
What techniques and methods are used?
The steps in this project are: 1. Growing up of bacteria containing the plasmid coding for the AAVLP HSP vaccine. 2. Purify the plasmid DNA from the bacteria. 3. Transient transfection of HEK293 cells using the PCI method. 4. Fast Protein Liquid Chromatography (FPLC) purification of the AAVLP HSP particles using an Âkta system. 5. Analyze the purified virus titer using a capsid specific enzyme-linked immunosorbent assay (Elisa), and polyacrylamide gel electrophoreses (PAGE).
What is the role of the student?
What are the tasks expected to be accomplished by the student?
A plasmid has been generated in the lab where the AAVLP protein VP3 of AAV virus is mutated at site 453 and 587 to contain part of the sequence coding for HSP70. The student will grow up the bacteria containing the plasmid. After the bacteria has been grown up, the plasmid will be isolated from the bacteria. After that the plasmid has been isolated, the plasmid will be transient transfected in HEK293 cells. After transfection the cells will be grown for 3 days in which time the AAVLP HSP particles are produced. The AAVLP HSP capsids will be purified from the HEK293 cells using a newly developed column material (AVB). This purification is performed by FPLC on the Äkta machine. To optimize the purification different loading and elution buffers will be tested. After purification the purity and titer of the capsids will be analyzed by ELISA and PAGE analysis.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
No theoretical teaching provided.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic understanding of molecular biology and protein chemistry.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system
- Development of AAVLP(HPV16/31L2) Particles as Broadly Protective HPV Vaccine Candidate. Karen Nieto; Margit Weghofer; Peter Sehr; Mirko Ritter; Sebastian Sedlmeier; Balasubramanyam Karanam; Hanna Seitz; Martin Muller; Markus Kellner; Markus Horer; Uwe Michaelis; Richard B. S. Roden; Lutz Gissmann; Jurgen A. Kleinschmidt. Nieto K; Weghofer M; Sehr P; Ritter M; Sedlmeier S; et al. (2012) Development of AAVLP(HPV16/31L2) Particles as Broadly Protective HPV Vaccine Candidate. PLoS ONE 7(6): e39741. doi:10.1371/journal.pone.0039741
- Mosenson J a; Zloza A; Nieland JD; Garrett-Mayer E; Eby JM; Huelsmann EJ; et al. Mutant HSP70 reverses autoimmune depigmentation in Vitiligo. Sci Transl Med [Internet]. 2013;5(174):174ra28. Available from: http://www.ncbi.nlm.nih.gov/pubmed/23447019
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