Biomedical Engineering Reference
In-Depth Information
Support
function
Coupling
agent
Active
intermediate
Activation
conditions
Coupling
conditions
Major reacting groups
on proteins
pH 11-12.5
2M carbonate
pH 9-10.
24 hr at 4
—NH
2
OH
CNBr
O
CN
H
°
C
O
OH
Cl
Cl
N
OH
O
Cl
N
N
N
Benzene
2 hr at 50
pH 8.
12 hr at 4
—NH
2
or
°
C
C
0.1M phosphate
°
N
N
R
NH
2
R Cl, NH
2
,
OCH
2
COOH, or
NHCH
2
COOH
R
S
10% thiophosgene
CHCl
3
,
reflux reaction
pH 9-10. 0.05M HCO
−
2 hr at 25
°
C
Cl
NH
2
Cl
C
N
C S
O
Same as isothiocyanate
Same as isothiocyanate
NH
2
Cl
C
Cl
N
C
O
—NH
2
O
O
O
H
C
2.5% Glutaraldehyde
in pH 7.0, 0.1M PO
4
pH 5-7, 0.05 M phosphate,
3 hr at R.T.
(CH
2
)
3
CH
NH
2
N
OH
HC(CH
2
)
3
CH
O
O
H
2
C
C
O
1% Succinic
anhydride, pH 6
See carboxyl
derivatives
(CH
2
)
2
C
O
NH
C
NH
2
OH
H
2
C
C
O
—NH
2
2N HCl: 0.2g NaNO
2
at 4
°
C for 30 min
(reaction conditions for
aryl amine function)
pH 8, 0.05M bicarbonate.
1-2 hr at 0
°
C
+
N
—SH
NH
2
N
HNO
2
OH
O
C
O
NH
2
pH 8, 0.05M bicarbonate.
1-2 hr at 0
°
C
H
2
N
NH
2
NH
2
—SH
C
N
3
HNO
2
OH
O
50mg 1-cyclohexyl-3-
(2-morpholinoethyl)-
carbodiimide metho-p-
toluene sulfate/10ml,
pH 4-5
2-3 hr at R.T.
pH 4,
2-3 hr at R.T.
R'
R'
NH
2
C
N
N
O
OH
C + H
+
N
C
C
or
NH
+
R
R
SH
or
(Intermediate formed
from carboxyl group are either
protein or matrix)
O
C
O
−
10% Thionyl chloride/CHCl
3
,
reflux for 4 hr
pH 8-9,
1 hr at R.T.
—NH
2
O
O
C
Cl
C
SOCl
2
OH
O
O
C
0.2% N-hydroxysuccinimide,
0.4% N,N-dicyclohexyl-
carbodiimide/dioxane
pH 5-9,
0.1M phosphate,
2-4 hr at 0
°
C
—NH
2
O
HO
N
C
O
N
C
C
OH
O
O
Fig. 3.2.16-2 Examples of various chemical methods used to bond biomolecules directly to reactive supports (
Carr and Bowers, 1980
).
