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Enzyme Purification by Size Exclusion (SEC) and Gel Filtration (GF)

Based on different characteristics of enzyme molecules, Creative Enzymes has established several methods for enzymes isolation and purification. Size exclusion chromatography (SEC) is based on the variation of relative molecular mass of different enzymes. Creative Enzymes has a skilled technical team to achieve efficient separation of enzyme mixtures. Creative Enzymes offers the best enzyme purification by size exclusion and gel filtration relying on the extensive experiences, a large reagent library, and excellent operation facilities.

Size exclusion chromatography (SEC) is also known as gel filtration, gel permeation, or sieve chromatography. In addition to separating proteins with different molecules sizes, SEC can be used to resolve oligomeric forms of a particular protein. Furthermore, SEC can be also used to exchange the buffer of a sample for a different one. In the SEC process, a porous gel, the sieving medium is needed. Molecules much smaller than the pore diameter will have more probability of penetrating the gel, so they will pass through the column more slowly and have relatively longer preserve time. Therefore, the large enzyme molecules will be first eluted and collected, before the small compounds such as ions and buffer salts.

Cutoff size, void volume, and column volume are three important parameters to characterize size exclusion columns. Cutoff size refers to the approximate size of the largest molecule that may enter the beads. The column volume refers to the total accessible volume of the column to solvent.

Enzyme Purification by Size Exclusion (SEC) and Gel Filtration (GF) Figure 1: Theoretical gel filtration chromatograph of a sample. In this case, protein which is larger than the cutoff size elutes at the void volume. The majority of the protein elutes within the separation range of the column and is separated from the aggregates. The protein sample is originally in a buffer of higher conductivity than that of the column and excess salts eluted at the column volume.

Enzyme Purification by Size Exclusion (SEC) and Gel Filtration (GF) Figure 2: The structure of dextran bead and the isolation process by SEC.

Creative Enzymes has plenty of experiences to customize the separation process for the target enzyme. We examine all factors that will influence the elution process, such as the bead density and the length of the column. As for enzyme purification by SEC, Creative Enzymes has reached a high level of proficiency and can perform purification in a rapid turnover as well as high quality. We sincerely look forward to the opportunity of presenting you the superior and professional services.

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