Naslov
Generation of ammodytoxin-anti-cathepsin B immuno-conjugate as a model for delivery of secretory phospholipase A2 into cancer cells

Autori
Premzl, Ales ; Kovačić, Lidija ; Halassy, Beata ; Križaj, Igor

Izvornik
Toxicon (0041-0101) 51 (2008), 5; 754-764

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
snake venom; Vipera ammodytes ammodytes; secreted phospholipase A2; ammodytoxin; immuno-conjugate; cancer

Sažetak
Ammodytoxin (AtxC) is a toxic secreted phospholipase A2 (sPLA2) from the venom of Vipera ammodytes snake. To evaluate its potential to kill cancer cells, the toxin was cross-linked to the monoclonal antibody against cathepsin B which endocytose upon binding to cathepsin B, an antigen overexpressed on the plasma membrane of cancer cells. Sulfo-SBED derivative of AtxC, possessing the same biological activity as the wt toxin, was photo-reacted with antibodies to form a covalent immuno-conjugate. In the cytosol-like conditions AtxC was gradually released from the conjugate due to reduction of the disulfide bond in the spacer arm of the cross-linker. The pshospholipase activity of the preparation reached maximum in 10 minutes and then decreased gradually. As demonstrated by fluorescence microcopy, the immuno-conjugate targeted Caco-2 colon adenocarcinoma cells but was just very slowly internalized, the likely reason of only slight citotoxicity observed in MTT test. Despite the lack of clear cytotoxic effect of AtxC-antibody conjugate od Caco-2 cells we demonstrated in this work a new methodology for targeted delivery of the (toxic) sPLA2 into cells, promising in research or therapy.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti, Drvna tehnologija

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