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J. Lipid Res.
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A more recent version of this article appeared on February 1, 2004

Papers In Press, published online ahead of print November 16, 2003
J. Lipid Res., doi:10.1194/jlr.D300023-JLR200
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Submitted on July 1, 2003
Revised on October 1, 2003
Accepted on November 5, 2003

Quantitation of the active and low-active forms of human plasma phospholipid transfer protein by ELISA

Sarah Siggins, Minna Kärkkäinen, Jukka Tenhunen, Jari Metso, Esa Tahvanainen, Vesa M Olkkonen, Matti Jauhiainen, and Christian Ehnholm

Department of Molecular Medicine, National Public Health Institute, Helsinki FIN-00251

Corresponding Author: sarah.siggins{at}ktl.fi

Human plasma contains two forms of PLTP, one catalytically active (HA-PLTP) and the other a low-active (LA-PLTP) form. We present here a PLTP ELISA that allows not only for accurate measurement of PLTP concentration in plasma but also of the distribution of both LA- and HA-PLTP. To achieve similar immunoreactivity of the two PLTP forms, a denaturing sample pre-treatment with 0.5 % SDS was required. Distribution of LA- and HA-PLTP in plasma was assessed using size-exclusion chromatography, Heparin-Sepharose chromatography, anti-PLTP immuno-affinity chromatography, and dextran sulfate-CaCl2 precipitation. All four methods demonstrated that approximately 60 % of plasma PLTP represents LA- and 40 % HA-PLTP. According to the modified ELISA, the total serum PLTP concentration in a random Finnish population sample (n = 80) was 5.81 ± 1.33 mg/l (mean ± S.D.) (range 2.78 - 10.06 mg/l) and the mean activity 5.84 ± 1.39 µmol/ml/h (range 3.21 - 11.15 µmol/ml/h). To quantitate both forms of PLTP in sera from this sample, we combined dextran sulfate-CaCl2 precipitation with the modified PLTP ELISA. The HA-PLTP mass (mean 1.87 ± 0.85 mg/l) correlated significantly with serum PLTP activity, while that of LA-PLTP (mean 3.94 ± 1.4 mg/l) showed no correlation with phospholipid transfer activity


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