Pathogenesis of Lung Cancer: Immunoaffinity-Proteomics
|Coordinator||Karolinska institutet - Institutionen för onkologi-patologi|
|Funding from Vinnova||SEK 1 999 821|
|Project duration||October 2010 - March 2014|
Purpose and goal
The original aim of the project is elucidating the mechanism of heterogonous lung cancer using proteomics approach. Since fresh samples were hardly collected, the plan has been changed for the establishment of comprehensive proteomics method for small amount of cell samples. A meter-long monolithic column was applied to conduct proteome from small amount of immune cells as a model of heterogeneous cancer tissue. 350 proteins were identified from 1000 cells by the method and further investment is expected, however, due to insufficient resource, instruments time was not secured.
Results and expected effects
Due to change of a situation, fresh samples from patients could not be obtained for the project. Thus, the project aimed to develop the method for epithelial or immune cell specific proteomics in heterogeneous cancer tissues. The workflow consisting of EpCAM (++) cell sorting by FACS, MS-friendly sample preparation, highly sensitive a meter-long monolithic column and an in-house mass spectrometry method was established. The workflow will be useful to investigate the role of immune cells in disease.
Approach and implementation
Problem-solving ability is an essential for successful completion of the project. The hardest problem in this project was frequent policy and priority changes of clinical sample collection. As a foreign researcher, some guide and help for collaborating with physicians are necessary, but it was not always expected. Also, securing the instrument time from a core facility, which is my host group, was difficult. These experiences suggest me to secure multiple groups to conduct research promptly as well as negotiating with several different milieu.