1. THE SCIENCE BEHIND MASS SPECTROMETRY (Biology of a Chemistry Machine) SARAVANAN KUMAR , Research Associate , Proteomics Facility, Plant Transformation Group, International Centre for Genetic Engineering & Biotechnology, New Delhi National Agricultural Innovation Project (NAIP) supported workshop on Proteomics: High Throughput Analysis of Proteins and Proteome by Mass Spectrometry” 22 nd - 26 th March 2010 ICGEB, New Delhi DR. V. SIVA REDDY , Organizer & Consortium Principal Investigator , NAIP component -4 , International Centre for Genetic Engineering & Biotechnology, New Delhi

2. Thanks to them

3. Mass Spectrometry

4. Mass Spectrometry/meter..... Mass spectrometry – Science – based on the motion of charged particles in a electric or magnetic field Motion- movement- mass/charge (m/z) z- if the charge is known, the mass of the ion is known m/z – m + or (m+H) + m/z= (M+n) /n M= (m/z)*n-n Transfers sample in to vacuum Ionization/ acceleration Flight/ resolution Signal detection

5. Inside the MS meter ,Ionisation.... ESI- Electro spray ionisation LDI –Laser desorption ionisation

6. MALDI - acceleration....

7. ESI - acceleration....

8. Electro spray & Laser desorption

13. Mass calculation ESI -MS m/z= (M+n) /n

14. m/z= (MW+n) /n m/z=(MW+n+1) / (n+1) m/z= (MW+n) /n

18. ESI-MS of peptide

19. ESI MS of Protein

20. MALDI MS of Protein 0.0 0.2 0.4 0.6 0.8 1.0 4 x10 Intens. [a.u.] 15000 20000 25000 30000 35000 40000 45000 50000 55000 m/z 26661.395 53321.02 13331.20

21. MALDI MS of Peptide Diff m/z= 22 da 2196.128 2218.106

23. Peptide sequence based on Backbone ions Peptide ion formation

24. PTM identification/prediction

26. For the Real time Experts (Bench workers) http://www.abrf.org/index.cfm/dm.home

30. LC-ESI MS

31. LC-MALDI -MS

35. You need to be aware of what others are doing, applaud their efforts, acknowledge their successes, and encourage them in their pursuits. When we all help one another, everybody wins.