Biomedical Engineering Reference
In-Depth Information
(conjugated) to carrier molecules. Such macromolecular carriers
are, for example, the hemolymph of the keyhole limpet
Megathura
crenulata
(KLH), the hemolymph of the eatable snail
Helix poma-
tia
, the hemolymph of the horseshoe crab
Limulus polyphemus
,
cationized serum albumin, hen egg ovalbumin, thyreoglobulin, or
dextranes.
To get a directed coupling of a peptide via its N-terminal NH
2
-
group, the introduction of a monochloroacetylglycyl residue during
peptide synthesis is recommended. Of course, coupling via NH
2
-,
SH-, or carboxyl groups of peptide side chains is also possible.
Production of antiserum with high titer and specifity is done
by trial and error, especially because each immunized animal gives
antisera with different characteristics; therefore, several groups of
animals should be immunized with different antigen preparations.
Universal haptens, suitable especially in immunohistochem-
istry, dot blots, Western blots, or other kinds of immunoassays,
are, for instance, biotin, fluorescein, or digoxigenin.
Some coupling reagents suitable for preparation of hapten-
carrier conjugates are given in Table 4.1.
Table 4.1.
Selected reagents for protein and peptide conjugation to
carrier proteins
Coupling reagent
Peptide Carrier protein
Binding to
1-Cyclohexyl-3-(2-morpholin-4-
-NH
2
, -COOH
-COOH, -NH
2
yl-ethyl) carbodiimide
1-Ethyl-3-(3-dimethyl-
-NH
2
, -COOH
-COOH, -NH
2
aminopropyl) carbodiimide
(EDC; EDAC)
2-Iminothiolane (Traut's
C lCH
2
CO-NH-CH
2
-CO-
-SH
Reagent)
3-Maleimidobenzoic acid (MBS)
-SH, -NH
2
-NH
2
,-SH
N-hydroxysuccinimide ester
Succinimidyl 4-(N-male-
imidomethyl)-cyclohexane-1-
carboxylate (SMCC)
-SH, -NH
2
-NH
2
,-SH
Aryldiazonium chloride
-SH, -Tyr
-SH, -Tyr
Bis-diazotated benzidine (BDB)
-Tyr
-Tyr
Glutardialdehyde
-NH
2
-NH
2
Sodium periodat
-NH
2
Saccharide
side chain
N-Succinimidyl-3-(2-pyridyl-
dithio) propionate (SPDP)
-NH
2
-NH
2
4-Azido-benzoic acid
N-hydroxysuccinimide ester
(HSAB)
(photoactivation)
-NH
2
