Reverse calcium affinity purification of Fab with calcium derivatized hydroxyapatite.
This study introduces the application of calcium-derivatized hydroxyapatite for purification of Fab. Fab binds to native hydroxyapatite but fails to bind to the calcium derivatized form. IgG, Fc, and most other protein contaminants bind to the calcium form. This supports Fab purification by a simple flow-through method that achieves greater than 95% purity from papain digests and mammalian cell culture supernatants. Alternatively, Fab can be concentrated on native hydroxyapatite then eluted selectively by conversion to the calcium-derivatized form.
Gagnon
Pete
P
Validated Biosystems, San Clemente, CA 92672, USA. pete_gagnon@mac.com
Cheung
Chia-Wei
CW
Yazaki
Paul J
PJ
D002118:Calcium / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D003470:Culture Media; D017886:Durapatite / Q000737:chemistry*; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000302:isolation & purification
D003470:Culture Media; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D017886:Durapatite; D002118:Calcium