BYU

Abstract by Andikan Nwosu

Personal Infomation


Presenter's Name

Andikan Nwosu

Degree Level

Doctorate

Co-Authors

Yongchenz Cong
Ryan Kelly

Abstract Infomation


Department

Chemistry and Biochemistry

Faculty Advisor

Ryan Kelly

Title

OPTIMIZED MASS SPECTROMETER-BASED WORKFLOW FOR SMALL AND TRACE FORMALIN FIXED PARAFFIN EMBEDDED SAMPLES USING THE NANOPOTS SYSTEM

Abstract

Formalin fixing followed by paraffin embedding (FFPE) is a common and cost-effective method for preserving tissues in clinical settings. These samples remain stable for long periods of time and are useful sources for biomedical research. However, retrieving proteins from FFPE tissues can be daunting due to the crosslinks formed after formalin fixation. Extracting proteome information by Mass spectrometry from these tissues has already been accomplished but only on a large scale which masks cell to cell heterogeneity. Here we present an MS-based workflow for nano scale FFPE samples containing as little as 30ng protein for proteomic analysis. We employed the use of laser cut microdissection to isolate individual tumor tissue cells and prepared samples in an inhouse fabricated nanochip using our custom nanoPOTS platform Preliminary MS analysis identified approximately 3000 peptides and 1200 proteins in 200um mouse kidney tissue xenograft. Our next line of action is to optimize our protein extraction conditions and apply them to Pancreatic ductal adenocarcinoma tissues. But our preliminary data already show great promise for the use of nanoPOTS with FFPE sample