Biomedical Engineering Reference
In-Depth Information
(a)
Molar coupling
ratio
Reactivity in cell-
based potency assay%
100
Unlabeled
0.33
100
0.66
100
1.4
74
2.3
42
(b)
Molar
coupling
Molar
incorporation
Biacore
relative binding
Affinity
kD (pM)
Label
ratio
ratio
Unlabeled
NA
NA
100%
579
Fluorochrome 1
10
2.3
93%
507
Fluorochrome 1
20
10.6
76%
747
Fluorochrome 2
4
1.1
85%
572
Fluorochrome 2
8
5.6
65%
1020
(c)
1:15
1:30
1:60
10 0
10 1
10 2
10 3
10 4
10 0
10 1
10 2
10 3
10 4
10 0
10 1
10 2
10 3
10 4
FL4-H
FL4-H
FL4-H
(d)
Lot
Molar coupling
Molar
incorporation
ratio
GMFI
ratio
Lot 1
15
6.2
825
Lot 2
15
6.6
1052
Lot 3
15
9.0
1182
Lot 4
15
6.6
1090
FIGURE 10.1 The effect of increasing the molar challenge ratio (label:biotherapeutic) used
during conjugation on biological activity (a), affinity to target (b), and cell binding profile (c).
Reproducibility of conjugation procedure was determined and measured by cell binding
intensity (d).
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