AN ELISA FOR TISSUE FACTOR USING MONOCLONAL-ANTIBODIES

Whereas tissue factor, a high-affinity cell-surface receptor and essential cofactor for the serine protease factor VII is constitutively present in certain tissues such as epithelial tissue, brain and placenta, it is not normally expressed by cells within the vasculature. However, the stimulation of monocytes and endothelial cells by a variety of inflammatory and immunological reactions results in the induction of cell surface tissue factor (TF) expression. TF is also expressed on tumour cells, and may play a role in tumour growth and metastasis formation. To examine the role of TF in these processes we developed monoclonal antibodies to human tissue factor apoprotein. The antibodies were characterized by neutralization of the procoagulant activity and by immunoblotting. With two of these monoclonal antibodies a sandwich ELISA was developed for the rapid quantitation of TF. The sensitivity of the assay permits extensive studies involving the modulation of TF expression on small numbers of cells. The results are comparable to the functional clotting assay as evaluated with unpurified TF and with the tumour cell line MCF-7. For certain applications, monitoring of cellular TF expression by ELISA using anti-TF monoclonal antibodies is preferable because it is not influenced by other coagulation factors or by inhibitors of procoagulant activity on the cells.