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Size Exclusion Chromatography (SEC) is a chromatographic method which separates analytes solely based on their size, where molecules are separated on the basis of their exclusion from pores in the column packing material. Larger analytes will elute first, while the smaller molecules interact more with the stationary phase and will elute later.
Based on the analytes and the mobile phases used, size exclusion chromatography can be broken into two main categories, aqueous size exclusion/gel filtration chromatography (GFC) and non-aqueous size exclusion/gel permeation chromatography (GPC).
Aqueous Size Exclusion/ Gel Filtration Chromatography (GFC)
Analyze and/or characterize proteins, peptides, and other biomolecules such as antibodies, immunoglobulins, protein complexes, protein aggregates, and desalting.
Non-Aqueous Size Exclusion/ Gel Permeation Chromatography (GPC)
Analyze polymers such as adhesives, oils, plasticizers, plastics, resins, and rubbers/elastomers.
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An introduction to the method development strategies of gel permeation chromatography.
GFC for protein aggregation and protein post-translational modifications (PTMs)
Gel filtration chromatography (GFC) is used to separate large macromolecules such as antibodies, immunoglobulins, protein complexes, protein aggregates, peptides, and other biomolecules. In gel filtration chromatography, the compounds of interest “filter” through the stationary phase based on their size.
Typically aqueous solvents are used in the mobile phase to ensure that the compound of interest maintains biological activity. Gel filtration columns can separate biomolecules that range from 200 to 1,500,000 Daltons in size. Gel filtration chromatography columns require a GFC column that has low surface activity, high efficiency, and consistent uniform pore size.
Method Development Strategies for Intact Protein Analysis by RP and GFC
Optimize HPLC|UHPLC and GFC separations for proteins, peptides, and other biomolecules.
The GPC Solvent Savings Calculator allows you to quickly calculate the solvent savings and cost savings you can achieve with your GPC polymer analysis.
Gel Permeation Chromatography (GPC) is an effective means of separating and analyzing polymers, resins, oils, plastics, plasticizers, and small organics whose molecular weight can vary from 100-10,000,000 Daltons. HPLC columns used for GPC are packed with polymeric media that can range in pore size from 50 Å to 10E6 Å and can be used with organic solvents such as toluene, THF, methanol, DMF, and methylene chloride.
Ideal gel permeation chromatography requires a GPC column that has superior mechanical strength and extreme solvent and temperature compatibility.
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GPC is principally used to separate polymers and organic molecules based on their size
Simple-to-use SecurityGuard GPC protects ANY non-aqueous GPC column. Extremely economical alternative to the typical GPC guard columns.
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