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Related Experiment Videos

New developments in biochemical mass spectrometry: electrospray ionization.

R D Smith1, J A Loo, C G Edmonds

  • 1Chemical Sciences Department, Pacific Northwest Laboratory, Richland, Washington 99352.

Analytical Chemistry
|May 1, 1990
PubMed
Summary

Electrospray ionization-mass spectrometry (ESI-MS) enables accurate molecular weight determination for large biological molecules like proteins. Advanced techniques like tandem mass spectrometry and hyphenation with separation methods offer powerful structural analysis capabilities for biomolecules.

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Biophysics

Background:

  • Electrospray ionization-mass spectrometry (ESI-MS) is a crucial technique for analyzing biological compounds.
  • Accurate molecular weight determination is essential for characterizing proteins and oligonucleotides.
  • Advancements in instrumentation have expanded the scope of ESI-MS applications.

Purpose of the Study:

  • To review the principles, development, and recent applications of ESI-MS for biological compounds.
  • To highlight the capabilities of ESI-MS in determining molecular weights of large biomolecules.
  • To discuss the potential for structural studies using ESI-MS and related techniques.

Main Methods:

  • Electrospray ionization-mass spectrometry (ESI-MS) for molecular weight determination.

Related Experiment Videos

  • Tandem mass spectrometry (MS/MS) with collision-induced dissociation (CID) for structural analysis.
  • Coupling ESI-MS with capillary electrophoresis (CE) and microcolumn liquid chromatography (LC) for on-line separation and analysis.

    • Main Results:

    • ESI-MS allows accurate molecular weight determination for proteins >50,000 Da, and even >100,000 Da.
    • Similar capabilities are being developed for oligonucleotides.
    • Detector sensitivity is correlated with molecular weight, while ionization efficiency remains high.
    • Effective CID is achievable for large molecules like albumin (approx. 66,000 Da).
    • On-line separation and analysis of subpicomole protein quantities are possible using CE-ESI-MS and LC-ESI-MS.

    Conclusions:

    • ESI-MS is a powerful tool for characterizing large biological molecules.
    • Tandem MS and hyphenated techniques significantly enhance structural elucidation capabilities.
    • Future developments promise even greater sensitivity and advanced structural information retrieval.