FIELD: chemistry.
SUBSTANCE: invention relates to biotechnology and discloses a tissue factor-bearing microvesicle. A tissue factor (TF)-bearing yeast derived microvesicle, which shortens coagulation time, contains (i) a yeast membrane and (ii) a human tissue factor (TF) protein or a variant thereof having pro-coagulant activity, wherein a portion of said human tissue factor (TF) protein having pro-coagulant activity is integrated in said membrane, and where the N-terminal domain of said human TF protein or version thereof having pro-coagulant activity lies on the outer side of said membrane, and where said variant thereof having pro-coagulant activity matches the human TF protein having at least one modification selected from a group consisting of: inclusion of a heterologous sequence which is suitable for alleviating separation or purification of the version, wherein said heterologous sequence is bound with the C-terminal or N-terminal domain; removal of a signal peptide which leads to a mature human TF protein; modifying the N-glycosylation site which corresponds to residues in positions 11-13 of the mature human TF protein, wherein said modification makes said site non-functional; modifying the N-glycosylation site which corresponds to residues in positions 124-126 in the mature human TF protein, wherein said modification makes said site non-functional; modifying the N-glycosylation site which corresponds to residues in positions 137-139 in the mature human TF protein, wherein said modification makes said site non-functional; modifying N-glycosylation sites which correspond to residues 124-126 and 11-13 in mature human TF protein, wherein said modification makes said site non-functional; modifying N-glycosylation sites which correspond to residues 124-126 and 137-139 in mature human TF protein, wherein said modification makes said site non-functional; modifying N-glycosylation sites corresponding to residues 124-126, 11-13 and 137-139 in mature human TF protein, wherein said modification makes said site non-functional; removing the whole domain or a portion thereof, responsible for binding with FVIIa; and introducing a mutation in the domain which is responsible for binding with FVIIa, which leads to significantly low affinity to FVIIa.
EFFECT: invention enables to obtain a microvesicle which shortens coagulation time.
16 cl, 34 dwg, 16 tbl, 6 ex
