A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms

The dimeric ectonucleotidase CD73 catalyzes the hydrolysis of AMP at the cell surface to form adenosine, a potent suppressor of the immune response. Blocking CD73 activity in the tumor microenvironment can have a beneficial effect on tumor eradication and is a promising approach for cancer therapy. Biparatopic antibodies binding different regions of CD73 may be a means to antagonize its enzymatic activity. A panel of biparatopic antibodies representing the pairwise combination of 11 parental monoclonal antibodies against CD73 was generated by Fab-arm exchange. Nine variants vastly exceeded the potency of their parental antibodies with ≥90% inhibition of activity and subnanomolar EC₅₀ values. Pairing the Fabs of parents with nonoverlapping epitopes was both sufficient and necessary whereas monovalent antibodies were poor inhibitors. Some parental antibodies yielded potent biparatopics with multiple partners, one of which (TB19) producing the most potent. The structure of the TB19 Fab with CD73 reveals that it blocks alignment of the N- and C-terminal CD73 domains necessary for catalysis. A separate structure of CD73 with a Fab (TB38) which complements TB19 in a particularly potent biparatopic shows its binding to a nonoverlapping site on the CD73 N-terminal domain. Structural modeling demonstrates a TB19/TB38 biparatopic antibody would be unable to bind the CD73 dimer in a bivalent manner, implicating crosslinking of separate CD73 dimers in its mechanism of action. This ability of a biparatopic antibody to both crosslink CD73 dimers and fix them in an inactive conformation thus represents a highly effective mechanism for the inhibition of CD73 activity.     

Selenicereus wittii (Cactaceae): An epiphyte adapted to Amazonian Igapó inundation forests

The biology, ecology, and distribution ofSelenicereus (Strophocactus)wittii, one of the least known taxa ofCactaceae, are described. This epiphyte climbs appressed to tree trunks with leaf-like, flattened stems and is found exclusively along the high waterline of black water rivers (Rio Negro, Vaupés, Apaporis) in the Igapó inundation forests of Amazonia. Ecophysiologically,S. wittii is a crassulacean acid metabolism (CAM) plant. It bears white, nocturnal flowers 25 cm in length which emit a fragrance consisting mainly of benzylalcohol, benzyl benzoate, and benzyl salicylate. They exhibit an extreme sphingophilous syndrome as an adaptation to pollination by probably only two species of hawkmoth from the generaAmphimoena andCocytius. The seeds, aberrant for the family, contain air-filled chambers and are water-dispersed. Thus,S. wittii represents the paradoxical life form of an hydrochorous epiphytic cactus which withstands periodical inundation.Dedicated to emer. Univ.-Prof. DrFriedrich Ehrendorfer on the occasion of his 70th birthday