Praktikum Practical course and Seminar Antibodies

Course Content: Participants in -Antibodies” obtain detailed knowledge of generating different antibody formats (polyclonal; monoclonal; recombinant; nanobodies) and methods using these antibodies in a variety of different cell biological and biochemical assays. During the scope of the practical course participants are introduced to antigen and antibody purification, ELISA, Co-Immunoprecipitation and Western blotting. Moreover, they are introduced to immofluorescence staining of fixed cells, as well as in vivo tracking of antigens using fluorescent nanobodies by confocal microscopy. During the course, the students will purify fluorescent proteins as potential new antigens via Ni-NTA columns as well as antibodies using protein G sepharose. Participants will perform an ELISA in a 96-well format to determine titer and subclass of polyclonal as well as monoclonal antibodies. In addition, students will analyse protein-protein interactions by Co-IP experiments of fluorescent fusion proteins, followed by Western blot analyses. Besides conventional antibody staining of fixed cells, participants will perform in vivotracking of antigens via nanobodies using confocal microscopy. A mandatory part of this course is an accompanying seminar, which entails the presentation of topics related to basic immunology, antibody formats and respective methods to support the practical course. Each student will have to present one topic. Emphasis is placed on hands-on practice with the techniques mentioned above and to specialized techniques. The lab entails 6 SWS, and requires a detailed lab report according to excellent scientific practice. Qualification goals: Students can apply theoretical and practical knowledge to approach biological questions in independent work. Students obtain skills for future lab work in the field of basic immunological methods with the focus on antigens and antibodies in particular in preparation for future lab work and master’s thesis. Students are well trained in good general lab practice, including standard safety procedures, precise handling of chemicals and instruments, conscientious documentation of lab procedures, and obtain skills in specialized techniques such antigen and antibody purification, ELISA, CO-IP, Western blotting and as well as confocal microscopy. Students practice critical evaluation and interpretation of data as a basis for careful and relevant conclusions. In working in small lab groups (2 students), social skills (teamwork, cooperation, fair play, work delegation, mutual respect), communication skills (rapport with instructors and fellow students, presentations, written lab reports), as well as organizational skills (efficient planning, documentation) are refined. Students learn excellent scientific methods in written data presentation, including well-founded introduction to the topic, documentation, interpretation and discussion of the results. These skills are particularly aimed at preparing students for thesis writing and scientific publications. Leonhardt, Heinrich , Prof.Dr. LMU München WiSe 1415 Dr. Meilinger Daniela