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JoVE Journal

Bioengineering

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Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow

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10:33 min

January 8th, 2014

January 8th, 2014

7,033 Views

0:05

Title

1:39

Preparation and Loading of Protein A Membrane Adsorber

3:36

Eluting, Concentrating, and Dialyzing the Protein

4:49

Quantify and Characterize Purified Product in an Expedited Western Blotting Procedure

7:00

Results: Characterization of Chimeric Murine Galectin-1 Fused to Human Fc (Gal-1hFc)

10:11

Conclusion

Compared with traditional affinity chromatography using protein A agarose bead-packed columns, protein A membrane adsorbers can significantly speed laboratory-scale isolation of antibodies and other Fc fragment-expressing proteins. Appropriate analysis and quantification methods can further accelerate protein processing, allowing isolation/characterization to be completed in one workday, instead of 20+ work hours.

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