Recombinant GPI-anchored HLA-A2.1 (HLA-A2.1-GPI/beta 2m) was used as a protein transfer vehicle to deliver a hepatitis B virus antigenic peptide to the surfaces of cytotoxic T cell targets. Empty HLA-A2.1-GPI/beta 2m was first produced in D. melanogaster cotransfectants and immunoaffinity purified. Cell coating with HLA-A2.1-GPI/beta 2m was shown to occur rapidly, and to be protein concentration dependent. Protein-transferred HLA-A2.1-GPI/beta 2m effectively presented a hepatitis B virus peptide to peptide-specific HLA-A2.1-restricted T cell clones in cytotoxicity assays. Protein transfer of functional GPI-modified class I MHC-antigenic peptide complexes represents a novel strategy for delivering functional antigenic complexes to cell surfaces that bypasses limitations of gene transfer and permits control of antigenic peptide densities at cell surfaces.