91
D *EtDSt3
o o E o E o E
1 +
E* I
(3)
Substituting these ratios with ratios expressed in terms of binding con
stants and ligand concentration gives,
D
D
o
+
D [L-K] [IJ [T7-M]
oKl=m i-0 Ki ^
1 +
m
ki
(6)
From liquid chromatography (Freeman, 1972),
D =
Ve Vra
(7)
where Ve, Vm and Vs are the elution volume, the volume of the mobile
phase and the volume of the stationary phase.
[I] [L-M]
_ D 1 + h D
Vs ~ D i + III D t + ill
Ki Kt
+ D,
[II[c-M]
KI^
i ill
Kt
(8)
A matrix based on Agarose would be sufficiently porous such that the con
centration of I and I would be equal, further the environment of the
stationary phase would be very much like the mobile phase in order to
justify the assumption, Kj = Kq. This simplifies equation (8) to.
Dq + Dq
[L^H]
kt^h
1+ifL
ki
[xjrc=Hi
kt
r t-r
i + -I1
(9)