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Clinical Chemistry 53: 1808-1813, 2007. First published August 23, 2007; 10.1373/clinchem.2007.089102
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(Clinical Chemistry. 2007;53:1808-1813.)
© 2007 American Association for Clinical Chemistry, Inc.

Lipids, Lipoproteins, and Cardiovascular Risk Factors

Development of a Novel Immunoassay for the Assessment of Plasma Gas6 Concentrations and Their Variation with Hormonal Status

Sylvain Clauser1,2,3, Séverine Peyrard4, Pascale Gaussem1,3, Malvina Crespin3, Joseph Emmerich3, Martine Aiach1,3 and Delphine Borgel1,2,a

1 Assistance Publique-Hôpitaux de Paris, Hôpital Européen Georges Pompidou, Service d’Hématologie Biologique A, Paris, France.
2 Université Paris-Sud, Laboratoire d’Hématologie, Orsay, France.
3 Institut National de la Santé et de la Recherche Médicale, Unité 765, Paris, France.
4 Assistance Publique-Hôpitaux de Paris, Hôpital Européen Georges Pompidou, Institut National de la Santé et de la Recherche Médicale, CIC 9201, Paris, France.

aAddress correspondence to this author at: Laboratoire d’Hématologie, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France. Fax 33 1 46 83 56 18; e-mail delphine.borgel{at}u-psud.fr.

Background: Gas6 is a vitamin K–dependent antiapoptotic protein that has been implicated in cardiovascular pathophysiology. We report the development and validation of an ELISA for Gas6, and the variation of plasma Gas6 with hormonal status in a study designed to evaluate the effect of oral contraception on plasma markers.

Methods: After validation of the main stages of the ELISA assay, we measured plasma Gas6 concentrations in 94 male and 88 female healthy volunteers ages 18 to 38 years. Forty-five of the women then received an oral contraceptive, which contained ethinylestradiol and levonorgestrel, for 3 months before a new measurement was performed at the same time point in their menstrual cycles.

Results: Interassay imprecision was 5.8%–11.8%, and the detection limit was 5.9 µg/L. Mean Gas6 plasma concentrations were significantly lower in men (52.0 µg/L) than in women not receiving oral contraceptives (63.8 µg/L, P <0.001). In the women who received oral contraceptives, Gas6 concentrations decreased after 3 months of therapy from 63.6 µg/L to 51.9 µg/L (P <0.001).

Conclusions: We have developed a simple and reproducible ELISA assay for measuring plasma Gas6 concentrations, which vary with sex and are decreased by oral contraceptive use. These results suggest regulation of plasma Gas6 concentrations by sex hormones. Future clinical studies may require participants to be stratified by sex.






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