Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Peptide characterization with a sulfoethyl aspartamide column.

D L Crimmins1, J Gorka, R S Thoma

  • 1Howard Hughes Medical Institute Laboratories, Washington University School of Medicine, St. Louis, MO 63110.

Journal of Chromatography
|June 29, 1988
PubMed
Summary

Strong cation-exchange (SCX) chromatography effectively separates peptides based on charge. This method enhances peptide characterization by resolving modified peptides that reversed-phase (RP) chromatography cannot, proving SCX

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

External validation of the Revised Cardiac Risk Index and National Surgical Quality Improvement Program Myocardial Infarction and Cardiac Arrest calculator in noncardiac vascular surgery.

British journal of anaesthesia·2019
Same author

Altered preoperative coagulation and fibrinolysis are associated with myocardial injury after non-cardiac surgery.

British journal of anaesthesia·2017
Same author

Leukotrienes biosynthesis in vascular surgery patients during perioperative period.

Journal of physiology and pharmacology : an official journal of the Polish Physiological Society·2014
Same author

The ABRF Edman Sequencing Research Group 2008 Study: investigation into homopolymeric amino acid N-terminal sequence tags and their effects on automated Edman degradation.

Journal of biomolecular techniques : JBT·2009
Same author

Nonlinear transmission of picosecond 10.6-Mu m pulses in InSb.

Optics letters·2009
Same author

Chemical cleavage of proteins on membranes.

Current protocols in protein science·2008

Area of Science:

  • Analytical Chemistry
  • Chromatography
  • Proteomics

Background:

  • Peptide analysis is crucial for understanding biological processes.
  • High-performance liquid chromatography (HPLC) is a standard technique for peptide separation.
  • Limitations exist in resolving modified or closely related peptide sequences using conventional methods.

Purpose of the Study:

  • To evaluate the utility of strong cation-exchange (SCX) HPLC for peptide characterization.
  • To compare SCX performance against standard C18 reversed-phase (RP) HPLC.
  • To demonstrate SCX's ability to resolve peptides based on charge and modifications.

Main Methods:

  • Analysis of over 50 peptides (5-20 residues) using a sulfoethyl aspartamide SCX column (200 x 4.6 mm).
  • A 60-min linear gradient elution (0-100% eluent B) at 1 ml/min was employed.

Related Experiment Videos

  • Comparison with a 60-min linear gradient RP-HPLC method using C18 column.

    • Main Results:

    • Peptide elution position correlated monotonically with the number of positively charged residues.
    • SCX successfully separated an arginine-rich peptide from its deleted counterpart, a feat not achieved by RP.
    • SCX and RP resolved methionine oxidation states differently, suggesting SCX's mixed-mode mechanism.
    • SCX differentiated sulfated and non-sulfated peptides, and N-terminally acetylated peptides from unmodified forms.

    Conclusions:

    • SCX-HPLC is a powerful tool for peptide characterization, offering superior resolution for charge-based separations.
    • SCX complements RP-HPLC by resolving peptides with specific modifications and charge variations.
    • The findings highlight SCX's utility in complex peptide mixture analysis and post-translational modification studies.