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The Journal of Nuclear Medicine Vol. 30 No. 8 1358-1366
© 1989 by Society of Nuclear Medicine
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In Vitro and In Vivo Targeting of Radiolabeled Monovalent and Divalent Haptens with Dual Specificity Monoclonal Antibody Conjugates: Enhanced Divalent Hapten Affinity for Cell-Bound Antibody Conjugate

Jean-Marc Le Doussal, Marie Martin, Emmanuel Gautherot, Michel Delaage and Jacques Barbet

Immunotech S.A., Campus de Luminy, Marseilles, France

Correspondence: For reprints contact: Jacques Barbet, Immunotech S.A., Luminy Case 915, 13288 Marseille Cedex 9, France.

ABSTRACT

A method of pretargeted immunolocalization of a mouse cell subset, using dual specificity monoclonal antibody conjugates and labeled divalent haptens, is described. Conjugates were prepared by coupling F(ab')2 fragments of an antibody specific for the allelic mouse B cell antigen Lyb8.2, to Fab' fragments of an anti-2,4-dinitrophenyl antibody. Divalent and monovalent haptens were obtained by coupling dinitrophenyl to peptides or to diethylenetriamine-pentaacetic acid. In vitro, divalent haptens bind with higher affinity to mouse spleen lymphocyte-bound than to excess soluble conjugate (affinity enhancement). In vivo, localization of 125I- or 111In-labeled divalent haptens in mouse spleen is much higher than that of the monovalent analogs. Thus, using divalent haptens, a new kind of specificity to target cells was achieved, suggesting that affinity enhancement may improve target to background ratios in radioimmunoscintigraphy.




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