Absolute size exclusion chromatography


Absolute size exclusion chromatography

Absolute Size Exclusion Chromatography (ASEC) is a technique which couples a dynamic light scattering (DLS) instrument to a size exclusion chromatography system for absolute size measurements of proteins and macromolecules as they elute from the chromatography system.

A measurement is said to be absolute if it doesn’t require calibration. In ASEC, there is no instrument or column to calibrate. The sizes of the macromolecules are measured as they elute in the flow cell of the DLS instrument.

Aside from the absence of costly and time consuming column calibration, a big advantage of DLS coupled with SEC is the ability to obtain enhanced DLS resolution. Batch DLS is simple and provides a direct measure of the protein size but the resolution of DLS is 3 to 1. Using SEC, the proteins and proteins oligomers are separated allowing oligomeric resolution. Aggregation studies can be done using ASEC.

In addition, size vs. mass relationships for globular proteins and polymeric systems are well known, and can be used to calculate the molecular mass from the DLS measured Stokes radius.

See also

* Dynamic Light Scattering
* Size Exclusion Chromatography


Wikimedia Foundation. 2010.

Look at other dictionaries:

  • Size-exclusion chromatography — Equipment for running size exclusion chromatography. The buffer is pumped through the column (right) by a computer controlled device Acronym SEC Classification Chromatography Analytes …   Wikipedia

  • Size exclusion chromatography — Infobox chemical analysis name = Size exclusion chromatography caption =Equipment for running size exclusion chromatography. The buffer is pumped through the column (right) by a computer controlled device acronym = SEC classification… …   Wikipedia

  • Absolute molar mass — is a process to determine the characteristics of molecules. TOC History The first absolute measurements (i.e. made without reference to standards) were based on fundamental physical characteristics and their relation to the molar mass. The most… …   Wikipedia

  • ASEC — may refer to: * ASEC Mimosas football club * ASEC Ndiambour football club * Absolute Size Exclusion Chromatography * Aviation Systems Engineering Company * Avenger Sebring Enthusiast Club * ASEC International …   Wikipedia

  • Multiangle light scattering — (MALS) is a technique for determining, independently, the absolute molar mass and the average size of particles in solution, by detecting how they scatter light. Collimated light from a laser source is most often used, in which case the technique …   Wikipedia

  • Molar mass distribution — In linear polymers the individual polymer chains rarely have exactly the same degree of polymerization and molar mass, and there is always a distribution around an average value. The molar mass distribution (or molecular weight distribution) in a …   Wikipedia

  • Inductively coupled plasma mass spectrometry — ICP MS Instrument Acronym ICP MS Classification Mass spectrometry Analytes atomic and polyatomic species in plasma, with exceptions; usually inte …   Wikipedia

  • Mark-Houwink equation — The Mark Houwink equation gives a relation between intrinsic viscosity [eta] and molecular weight M: [Paul, Hiemenz C., and Lodge P. Timothy. Polymer Chemistry. Second ed. Boca Raton: CRC P, 2007. 336, 338 339.] : [eta] =KM^aFrom this equation… …   Wikipedia

  • Mark–Houwink equation — The Mark–Houwink equation gives a relation between intrinsic viscosity [η] and molecular weight M:[1] [η] = KMa From this equation the molecular weight of a polymer can be determined from data on the intrinsic viscosity and vice versa. The values …   Wikipedia

  • Gel-Permeations-Chromatographie — Bei der Gel Permeations Chromatographie (GPC) handelt es sich um eine Art der Flüssigchromatographie ähnlich der HPLC. Die Trennung findet hier jedoch rein aufgrund der Größe (genauer: dem hydrodynamischen Volumen) der Moleküle in Lösung statt.… …   Deutsch Wikipedia