GE HEALTHCARE Biacore T100, Руководство пользователя

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11 Affinity in solution
11.2 Requirements for affinity in solution
204
Biacore T100 Software Handbook BR-1006-48 Edition AE
Rearranging gives
or
Solving for AB:
Substituting in the relationship B
free
= B
tot
– AB gives
This equation can be fitted to a plot of B
free
against A
tot
to calculate a value for
K
D
. (Formally, the equation has two solutions, but one is always negative and is
not meaningful in the context of an affinity determination.)
11.2 Requirements for affinity in solution
Affinity in solution experiments are run using a method. The method must be
correctly constructed as described in Section 5.10.4: if necessary, the keyword
table can be edited so that the conditions are met in full (see Section 6.6). Note
however that the command type cannot be edited in the keyword table. Refer
to Chapter 5 for details of how to construct methods in Method Builder.
Determination of affinity in solution is not supported by a wizard.
K
D
AB
⋅
A
tot
B
tot
AB A
tot
B
tot
+
( )
–
AB
2
+
⋅
=
AB
2
AB A
tot
B
tot
K
D
+ +
( )
–
A
tot
B
tot
⋅
+
0
=
AB
A
tot
B
tot
K
D
+ +
( )
2
-------------------------------------------
A
tot
B
tot
K
D
+ +
( )
2
4
----------------------------------------------
A
tot
B
tot
⋅
–
±
=
B
free
B
tot
A
tot
–
K
D
–
( )
2
-------------------------------------------
A
tot
B
tot
K
D
+ +
( )
2
4
----------------------------------------------
A
tot
B
tot
⋅
–
±
=