If an antibody must be purified from a small sample volume (0.5-10 mL), then the Amicon® Pro Purification System is recommended. Because this centrifugal device enables purification, buffer exchange, and concentration to be performed without transferring the sample or diluting it, the risk of antibody loss is minimized. The following method was developed for affinity purification of antibodies (0.5 mL sample) using protein A/G agarose resins in the Amicon® Pro System.
IgGs from rabbit serum (Cat. No. S20-100ML) were purified on Protein A or Protein G agarose, using either the Amicon® Pro System (10kDa MWCO, Cat Nos. ACK5010PA and ACK5010PG) or traditional spin columns. Purified IgGs were then buffer-exchanged and concentrated, either in the same Amicon® Pro devices or transferred from spin columns to separate Amicon® Ultra devices.
When the Amicon® Pro System was used, the process required 6 pipetting steps/spins, as compared to 11 pipetting steps/spins required for traditional spin columns and concentrators. As shown in the figure below, antibody recovery remained unchanged.
Antibody purification from rabbit serum using Amicon® Pro System and traditional spin columns (n=3)
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