Affinity Chromatography - Department of Molecular and Cellular ...

Sepharose
NH
(CH 2)
n
NH
N
N
O
N
N
HO
P
O
CH2
O
O
HO
O
HO
P
O
O
Fig. 43. Partial structure of 2'5' ADP Sepharose 4B.
NADP + -dependent dehydrogenases are also members of a larger group of proteins that
will interact with Procion Red, a synthetic polycyclic dye that shows certain structural
similarities to naturally occurring NADP + . When used as an affinity ligand attached to
Sepharose CL-6B, Procion Red HE-3B will bind strongly and specifically to a wide range of
proteins. Some proteins bind specifically due to their requirement for nucleotide cofactors,
while others, such as albumin, lipoproteins, blood coagulation factors and interferon, bind
in a less specific manner by electrostatic and/or hydrophobic interactions with the aromatic
anionic ligand.
NaO 3 S
SO 3 Na
NaO 3 S
N
N
N
N
SO 3 Na
OH
OH
SO 3 Na
NH
N
N
NH
SO 3 Na
N
NH
NH
N
O
N
N
OH
Sepharose
Fig. 44. Partial structure of Red Sepharose CL-6B.
75