In order to explore pathologies possibly associated with vitamin K deficiency, several monoclonal antibodies (mAbs) were produced against human Desgamma-Carboxy-Prothrombin (DCP). One of these mAbs, designated C4B6, detected DCP forms in the presence of Calcium ions, confirmed by comparison with the patterns of two electrophoretic techniques: Affino-Immuno-Electrophoresis (CAIE) and Polyacrylamide Gel Electrophoresis followed by Electro-blotting (PAGE-Blot). An Enzyme-Linked-Imunosorbent Assay (ELISA) using mAb C4B6 has been developed, optimized and standardized. It has proven to be specific for DCP forms and has a minimum sensitivity of 0.156 A.U/ml.

In the absence of vitamin K or in the presence of the vitamin K antagonists, abnormal nonfunctional forms of prothrombin circulate in the blood. A reliable and reproducible technique, derived from traditional crossed affinoimmunoelectrophoresis in presence of calcium lactate, was developed and optimized. The technique is based on nondenaturing polyacrylamide gel affinoelectrophoresis, with calcium lactate, of plasma samples, followed by immunoblotting with rabbit anti-human prothrombin serum and detection with an anti-rabbit immunoglobulin peroxidase conjugate. Depending on the plasmas, one or two bands were visualized and quantified by densitometry of the immunoblots. The technique was able to detect abnormal des-gamma-carboxylated prothrombins at concentration of 0.1 microgram/mL.