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Application of 2-D free-flow electrophoresis/RP-HPLC for proteomic analysis of human plasma depleted of multi high-abundance proteins by Robert L. Moritz; Andrew B. Clippingdale; Eugene A. Kapp; James S. Eddes; Hong Ji; Sam Gilbert; Lisa M. Connolly; Richard J. Simpson is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
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## Abstract Free‐flow electrophoresis (FFE) and rapid (6 min) RP‐HPLC was used to fractionate human citrate‐treated plasma. Prior to analysis, the six most abundant proteins in plasma were removed by immunoaffinity chromatography; both depleted plasma and the fraction containing the six abundant proteins depleted were taken for MS‐based analysis. Fractionated proteins were digested with trypsin and the generated peptides were subjected to MS‐based peptide sequencing. To date, 78 plasma proteins have been unambiguously identified by manual validation from 16% (15/96 FFE total fractions) of the collected FFE pools; 55 identifications were based on ≥2 tryptic peptides and 23 using single peptides. The molecular weight range of proteins and peptides isolated by this method ranged from ̃190 K (__e.g.__, Complement C3 and C4) to ̃4–6 K (__e.g.__, CRISPP and Apolipoprotein C1). This FFE/RP‐HPLC approach reveals low‐abundance proteins and peptides (__e.g.__, L‐Selectin ̃17 ng/mL and the cancer‐associated SCM‐recognition, immunodefense suppression, and serine protease protection peptide (CRISPP) at ̃0.5–1 ng/mL), where CRISPP was found in association with α‐1‐antitrypsin as a non‐covale
Who reads Application of 2-D free-flow electrophoresis/RP-HPLC for proteomic analysis of human plasma depleted of multi high-abundance proteins?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Robert L. Moritz; Andrew B. Clippingdale; Eugene A. Kapp; James S. Eddes; Hong Ji; Sam Gilbert; Lisa M. Connolly; Richard J. Simpson
- Publisher
- John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley; Research Square (ISSN 1615-9853)
- Published
- 2005
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)