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Monoclonal antibodies were raised against the surface of trypomastigote forms of Trypanosoma cruzi. Although some of these antibodies reacted against antigens shared by trypomastigote and epimastigote or amastigote forms, the majority were trypomastigote-specific. Trypomastigote-specific monoclonal antibodies recognized all infective stages, including trypomastigotes from the bloodstream of infected mice, insect feces, tissue culture and those resulting from differentiation of epimastigotes in axenic culture media. The monoclonal antibodies H1A10 and 6A2, as well as Fab fragments from H1A10, partially prevented T. cruzi invasion of LLC-MK 2 cell monolayers (inhibition of 50–70%) when present throughout the entire experiment. Both antibodies recognized an 85 kDa glycoprotein (Tc-85) of the trypomastigote surface which contains N-acetyl-d-glucosamine and/or sialic acid.