Program

Poster Presentations

Day 3, May 17（Thu.）　　Poster

3P-05　　PDF

Proteomic Analysis for a Slight Amount of Cancer Cells by iPP (in PCR plate) method

(¹Keio Univ., ²Keio Univ)
^oMasaru Mori^1,2, Sumiko Ohnuma¹, Masaru Tomita^1,2

We developed iPP (in-PCR tube) method, in which all steps of sample preparation in PCR tube and the reactive solution is directly injected from tube to LC/MS system, as a very simple and effective for proteomic analysis of a slight amount of cancer cells. The method was compared with iST (in-Stage tips) method known as a sample preparation for trace level proteomics. Five µL of HeLa cell lysate containing about 20 ng protein, which is expected from 100 HeLa cells, was subjected to reductive alkylation by DTT and IAA, digestion by trypsin, acidification by acetic acid to stop reaction in PCR tube or Stage tip. In addition, the desalting, elution, drying and dissolution was done in iST method. The samples were analyzed by LC/MS, and 152±38 and 141±41 proteins (from 476±164 and 476±198 peptide ions) was identified by iPP and iST method respectively. There is no significant difference between two methods in identification, but iPP method have simpler workflow keeping data qualities. So, iPP method is a good method for trace level proteomics.