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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2006;26:423.)
© 2006 American Heart Association, Inc.

Thrombosis

Development of ELISAs Measuring the Extent of TAFI Activation

Erik Ceresa; Els Brouwers; Miet Peeters; Christina Jern; Paul J. Declerck; Ann Gils

From the Laboratory for Pharmaceutical Biology and Phytopharmacology (E.C., E.B., M.P., P.J.D., A.G.), Faculty of Pharmaceutical Sciences, Katholieke Universiteit Leuven, Belgium; and The Institute of Clinical Neuroscience (C.J.), Sahlgrenska Academy, Göteborg University, Göteborg, Sweden.

Correspondence to Ann Gils, Laboratory for Pharmaceutical Biology and Phytopharmacology, Faculty of Pharmaceutical Sciences, Katholieke Universiteit Leuven, O&N2-PB824, Herestraat 49, B-3000 Leuven, Belgium. E-mail Ann.Gils{at}pharm.kuleuven.be

Objectives— To date, quantitation of TAFI antigen levels has been mainly focused on "total" antigen levels and has been shown to yield ambiguous results because of the existence of different isoforms and various degrees of activation. Our objective was to develop assays that allow measuring the extent of TAFI activation.

Methods and Results— A variety of enzyme-linked immunosorbent assays (ELISAs) were evaluated for their preferential reactivity toward TAFI before and after activation, and toward the recombinantly expressed activation peptide. Three ELISAs with distinct reactivities were selected: recognizing either exclusively nonactivated TAFI, the released activation peptide, or exclusively TAFIa (activated TAFI). Evaluation of TAFI activation during clot lysis revealed that decreases of TAFI levels are associated with increases of the released activation peptide and TAFIa levels. In addition, antigenic measurement of TAFIa parallels activity measured by chromogenic assay. Analyzing plasma samples revealed that subjects with hyperlipidemia had significantly higher plasma levels of both the activation peptide (109.2 versus 95.5; P<0.001) and TAFIa (112.1 versus 103.3; P=0.03), and not of TAFI antigen (92.5 versus 87.9; P=0.07) (results in % of plasma pooled from normolipidemic subjects).

Conclusion— ELISAs that allow to measure the extent of TAFI activation were developed. These ELISAs constitute more sensitive markers in studies on the relationship between TAFI and cardiovascular diseases.

We developed immunologic assays that allow to measure the extent of TAFI activation. These ELISAs constitute more sensitive markers in studies on the relationship between TAFI and cardiovascular related diseases.

Key Words: fibrinolysis • monoclonal antibodies • TAFI • ELISA • extent of TAFI activation

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