Performing a Purification of IgG Antibodies with rProtein A GraviTrap™ and rProtein A/Protein G GraviTrap™

Extracted from Affinity Chromatography Vol. 1: Antibodies, GE Healthcare, 2016
 

rProtein A GraviTrap™ are gravity-flow columns prepacked with 1 ml of rProtein A Sepharose® 4 Fast Flow. The columns are designed for fast and efficient manual purification of monoclonal and polyclonal antibodies, antibody fragments from cell culture supernatant, and biological fluids. The antibodies are simply captured with high specificity on protein A ligands in the gravity-flow columns. You do not need any other instrument with this protocol because the entire process relies on the flow of gravity. Together with the package that can be converted into a column stand (Workmate), parallel purification is made simple (see Figure 3.4). The columns are reusable up to five times.

Protein G GraviTrap

Figure 3.4. Protein G GraviTrap™ are prepacked gravity-flow columns that provide simple, manual purification of antibodies from cell culture.

 

A mix of rProtein A Separose® 4 Fast Flow and Protein G Sepharose® 4 Fast Flow is also available in GraviTrap™ format. The simple four-step procedure for purification using these GraviTrap™ columns is shown in Figure 3.5.

four-step procedure for purification using Protein G GraviTrap

Figure 3.5. Purifying antibodies with Protein G GraviTrap™ is a simple four-step procedure.

 

Sample preparation, buffer preparation, and purification

rProtein A GraviTrap™ as well as rProtein A/Protein G GraviTrap™ use the same protocol as Protein G GraviTrap.

Materials

     
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