P152-M Mass Spectrometric Identification of Proteins of Purified Protein Derivative (PPD) of Mycobacterium Bovis Stain AN-5 Using a 2D HPLC Protein Fractionation System

Tuberculin PPD Bovis, Intradermic (purified protein derivative of Mycobacterium bovis strain AN-5) was widely used for the Tuberculin test in cattle. However, protein components of PPD remain unknown since it is extremely difficult to separate PPD proteins by traditional protein fractionation techniques. A pilot mass spectrometric protein identification of PPD using the new protein fractionation system of Beckmen Coulter’s ProteomeLab PF 2D is reported here. ProteomeLab PF 2D is a two-dimensional liquid chromatographic system to fractionate complex proteomes. This system consists of a chromatofocusing column to separate the protein based on charge/pI in the first dimension and a reversed-phase column to separate the protein based on hydrophobicity in the second dimension. The advantages of this technique include high loading capacity and better detection of hydrophobic and low molecular weight proteins. Up to 800 protein fractions are collected. So far, 50 proteins such as cfp10, MPB 53, MPB 70, and others have been identified using MALDI-TOF or FT-ICR MS with Sequest or Mascot search engines. This study has demonstrate that ProteomeLab PF 2D with mass spectrometry is a potentially powerful tool to identify proteins from a large proteome.