Antibody Purification: Affinity Chromatography – Protein A and Protein G Sepharose

Affinity chromatography relies on the reversible interaction between a protein and a specific ligand immobilized in a chromatographic matrix

Ana Cristina Grodzki; Elsa Berenstein

2009

Scholarcy highlights

  • Affinity chromatography relies on the reversible interaction between a protein and a specific ligand immobilized in a chromatographic matrix
  • The sample is applied under conditions that favor specific binding to the ligand as the result of electrostatic and hydrophobic interactions, van der Waals’ forces and/or hydrogen bonding
  • After washing away the unbound material the bound protein is recovered by changing the buffer conditions to those that favor desorption
  • Methods are described for the purification of immunoglobulins, namely IgG, IgG fragments and subclasses, using the high affinity of protein A and protein G coupled to agarose
  • In the Subheading 3 there are protocols for affinity purification using a specific ligand coupled to commercial matrices like CNBr- Sepharose 4-B and Affigel

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