A wide range of protein isoforms in serum and plasma uncovered by a quantitative intact protein analysis system

@article{citeulike:2891720, abstract = {We have implemented an orthogonal 3-D intact protein analysis system (IPAS) to quantitatively profile protein differences between human serum and plasma. Reference specimens consisting of pooled Caucasian-American serum, citrate-anticoagulated plasma, and EDTA-anticoagulated plasma were each depleted of six highly abundant proteins, concentrated, and labeled with a different Cy dye (Cy5, Cy3, or Cy2). A mixture consisting of each of the labeled samples was subjected to three dimensions of separation based on charge, hydrophobicity, and molecular mass. Differences in the abundance of proteins between each of the three samples were determined. More than 5000 bands were found to have greater than two-fold difference in intensity between any pair of labeled specimens by quantitative imaging. As expected, some of the differences in band intensities between serum and plasma were attributable to proteins related to coagulation. Interestingly, many proteins were identified in multiple fractions, each exhibiting different pI, hydrophobicity, or molecular mass. This is likely reflective of the expression of different protein isoforms or specific protein cleavage products, as illustrated by complement component 3 precursor and clusterin. IPAS provides a high resolution, high sensitivity, and quantitative approach for the analysis of serum and plasma proteins, and allows assessment of PTMs as a potential source of biomarkers.}, address = {Department of Pediatrics, University of Michigan Medical Center, Ann Arbor, MI, USA; Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, MI, USA; PRS Proteomics, Ann Arbor, MI, USA; Department of Biological Chemistry, University of Michigan Medical Center, Ann Arbor, MI, USA; Fred Hutchinson Cancer Research Center, Seattle, WA, USA}, author = {David e and Kuick, Rork and Wang, Hong and Galchev, Vladimir and Deng, Bin and Zhao, Rong and Tra, John and Michael r and Amunugama, Ravi and Allen, David and Angela k and John r and Andrews, Philip and Gilbert s and Samir m}, citeulike-article-id = {2891720}, doi = {http://dx.doi.org/10.1002/pmic.200500103}, journal = {PROTEOMICS}, number = {13}, pages = {3343--3352}, posted-at = {2008-06-13 17:14:13}, priority = {2}, title = {A wide range of protein isoforms in serum and plasma uncovered by a quantitative intact protein analysis system}, url = {http://dx.doi.org/10.1002/pmic.200500103}, volume = {5}, year = {2005} }

TY - JOUR ID - citeulike:2891720 L3 - citeulike-article-id:2891720 TI - A wide range of protein isoforms in serum and plasma uncovered by a quantitative intact protein analysis system JF - PROTEOMICS VL - 5 IS - 13 SP - 3343 EP - 3352 AD - Department of Pediatrics, University of Michigan Medical Center, Ann Arbor, MI, USA; Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, MI, USA; PRS Proteomics, Ann Arbor, MI, USA; Department of Biological Chemistry, University of Michigan Medical Center, Ann Arbor, MI, USA; Fred Hutchinson Cancer Research Center, Seattle, WA, USA N2 - We have implemented an orthogonal 3-D intact protein analysis system (IPAS) to quantitatively profile protein differences between human serum and plasma. Reference specimens consisting of pooled Caucasian-American serum, citrate-anticoagulated plasma, and EDTA-anticoagulated plasma were each depleted of six highly abundant proteins, concentrated, and labeled with a different Cy dye (Cy5, Cy3, or Cy2). A mixture consisting of each of the labeled samples was subjected to three dimensions of separation based on charge, hydrophobicity, and molecular mass. Differences in the abundance of proteins between each of the three samples were determined. More than 5000 bands were found to have greater than two-fold difference in intensity between any pair of labeled specimens by quantitative imaging. As expected, some of the differences in band intensities between serum and plasma were attributable to proteins related to coagulation. Interestingly, many proteins were identified in multiple fractions, each exhibiting different pI, hydrophobicity, or molecular mass. This is likely reflective of the expression of different protein isoforms or specific protein cleavage products, as illustrated by complement component 3 precursor and clusterin. IPAS provides a high resolution, high sensitivity, and quantitative approach for the analysis of serum and plasma proteins, and allows assessment of PTMs as a potential source of biomarkers. AU - David e AU - Kuick, Rork AU - Wang, Hong AU - Galchev, Vladimir AU - Deng, Bin AU - Zhao, Rong AU - Tra, John AU - Michael r AU - Amunugama, Ravi AU - Allen, David AU - Angela k AU - John r AU - Andrews, Philip AU - Gilbert s AU - Samir m PY - 2005/// UR - http://dx.doi.org/10.1002/pmic.200500103 ER -