Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE credits
About 25% of the population in the industrialized countries suffers from allergic diseases. In Sweden, about 1/3 of the population in the age 25-45 suffer from allergy of which most common allergic are caused by pollen and fur. Today, there are several ways to treat allergens. The only treatment to obtain a long lasting effect is called allergen-specific immunotherapy. However, there are still difficulties to overcome with these therapies, including the risk of side effects and the risk to induce novel sensitization to proteins from the therapy.
The main wim with this project was to improve allergen specific immunotherapy by targeting effector cells, whoch can cause the symptoms, and the goal was to find binders for inhibitory receptors on mast cells and basophils tp down-regulate degranulation.
In this project I have therefore selected four affibodies using phage display that binds to two different inhibitory receptors expressed on mast cells and basophils. I have produced different constructs with these affibodies, and the original affibody, using a novel cloning system that has been developed during this time. The products contained a His6 tag for convenient purification using liquid chromotography (ÄKTA®) or magnetic beads. Finally, tje bomdomg am affinity of the produced constructs were analyzed ex vivo and in vitro using Flow cytometry (FACS), and Biacore®, respectively.
Preliminary results indicate that one of the selected affibodies bound the target, however more experiments are required to validate this finding.