Conjugation of enzymes

Protocol for the Two-Step Glutaraldehyde Conjugation of Enzymes to (Strept)avidin... 

Weston, P.D., Devries, J.A., and Wrigglesworth, R. (1980) Conjugation of enzymes to immunoglobulins using dimaleimides. Biochim. Biophys. Acta 612, 40-49. 

Conjugation of enzymes to antigens or antibodies were first developed for histochemical techniques and were used for localization of antigens and antibodies in tissue sections (60). 

Most popular and widely used procedures for the conjugation of enzymes to macromol-... 

OPDM gives superior results with BGase (100% conjugation of enzymes stable for at least 1 year at 4 C and pH 6.5). However, with APase 20-40% of the enzyme activity is lost. OPDM can be used for conjugation of almost all proteins. 

Some effective cross-linking agents are less suitable for the conjugation of enzyme to antibody. Diazonium salts of aromatic compounds react preferentially with histidine and tyrosine which are very often located in or near the active site of the enzyme. Protecting the active sites with inhibitors (Jansen et al., 1971) is often cumbersome and, consequently, not very popular. 

Fig. 11.12. Conjugation of enzyme to antibodies with m-maleimidobenzoyl-Af-hydrox-ysuccinimide ester (MBS) as described in Section 11.2.3.2.1.1.

Fig. 11.13. Conjugation of enzyme (1) to thiolated antibody (2) by 4- /V-maleimi-domethyl)-cyclohexane-l-carboxylic acid A-hydroxysuccinimide ester.

Conjugation of enzymes to Fab or IgG using the A -hydroxysuccinimide ester of 4-(A -maleimidomethyl)cyclohexane-l-carboxylic acid (CHM-NHS)... 

Conjugation of enzyme to protein A or IgG using A-succinimidyl-3-(2-pyridyldithio)-... 

M. J. Poznansky. Conjugates of enzymes and albumin immobilized proteins. Meth. Enzymol. 737 566(1988). 

T. Konno, J. Watanabe, K. Ishihara, Conjugation of enzymes on polymer nanoparticles covered with phosphorylcholine groups. Biomacromolecules 5 (2) (2004) 342-347. 

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