Maleimide Derivative of Hapten for Enzyme Coupling in EIA

  • Nobuo Monji
  • Albert Castro


For labeling of antibodies or macromolecular antigens with various enzymes, glutaraldehyde has been most widely used. In the case of β-galactosidase, maleimide derivatives have been used for coupling the enzyme to proteins, such as immunoglobulins and their fragments (Ishikawa, 1983). Use of these coupling agents showed no appreciable reduction of enzyme activity due to the involvement of sulfhydryl groups of the enzyme in the coupling process. Coupling of haptens to the enzyme, on the other hand, is often identical to the preparation of hapten-protein conjugates for immunization. Such coupling involves either amino or carboxyl groups of the enzyme, resulting in either low efficiency of coupling (Dray et al., 1975) or reduction of enzyme activity (Comoglio and Celada, 1976). In order to obivate such disadvantages, we utilized m-maleimidobenzoyl derivative of a hapten in order to couple it to sulfhydryl groups of the enzyme; a high efficiency of binding to the enzyme without appreciable reduction in enzyme activity were found.


Thin Layer Chromatography Sulfhydryl Group Maleic Anhydride Succinic Anhydride Appreciable Reduction 
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Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Nobuo Monji
    • 1
  • Albert Castro
    • 2
  1. 1.Genetic Systems CorporationSeattleUSA
  2. 2.Department of PathologyUniversity of Miami School of MedicineMiamiUSA

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