Biomedical Engineering Reference
In-Depth Information
Further coupling reagents are substances used in cross-linking
proteins, which allow an estimation of interacting area between
proteins or parts of a protein. Because of the chemical structure
of these substances, spacers are introduced between hapten and
carrier.
Cross-linking reagents may be divided into several types:
-
Water soluble (contain mostly sulfonic acid residues) and water
insoluble (penetrate mostly biological membranes)
-
Homobifunctional (two identically reactive groups) and heter-
obifunctional (at least two chemically different reactive groups)
-
Cleavable and not cleavable
Examples of cross-linking bifunctional agents are given in Table 4.2.
Table4.2.
Selectedhomo- andheterobifunctional cross-linkingreagents
Reagent
Homo
Hetero
Water
Cleavable
Distance
soluble
(nm)
Glutaraldehyde
x
Yes
No
Dimethyladipinimidate
(DMA)
x
Yes
No
0.86
Disuccinimidylsuberate
(DSS)
x
No
No
1.14
β
Bis[
-(4-azidosalicyl-
amido)-ethyl]-disulfide
(BASED)
x
No
Yes
3.47
Bis[2-(Sulfosuccinimido-
oxycarbonyl-oxy)ethyl]
sulfone (Sulfo-
BSOCOES)
x
Yes
Yes
1.30
Disuccinimidyltartrate
(DST)
x
Yes
Yes
0.64
Dimethyl-3,3
-
dithiobispropionimidate
hydrochloride (DTBP)
x
Yes
Yes
1.19
Succinimidyl 4-(N-
maleimidomethyl)-
cyclohexane-1-
carboxylate (SMCC)
x
o
o
1.16
m-Maleimidobenzyl-N-
hydroxysuccinimide
ester (MBS)
x
o
o
0.99
N-Sucinimidyl-3-(2-
pyridyldithio) propi-
onate (SPDP)
x
No
Yes
0.68
4-Sucinimidyl-4-(p-
maleinimidophenyl)
butyrat (SMPT)
x
No
Yes
1.12
