Quantitation of the active and low-active forms of human plasma phospholipid transfer protein by ELISA
| Autor: | Jari Metso, Vesa M. Olkkonen, Sarah Siggins, Christian Ehnholm, Matti Jauhiainen, Esa Tahvanainen, Minna Kärkkäinen, Jukka Tenhunen |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Adult
Protein Denaturation Phospholipid Enzyme-Linked Immunosorbent Assay QD415-436 Sensitivity and Specificity Biochemistry law.invention chemistry.chemical_compound Endocrinology Finnish population Affinity chromatography law Phospholipid transfer protein Humans Phospholipid Transfer Proteins Aged Chromatography low-activity form of phospholipid transfer protein Antibodies Monoclonal Membrane Proteins Reproducibility of Results lipid transfer protein Blood Proteins Cell Biology Middle Aged Reference Standards Blood proteins Recombinant Proteins high-activity form of phospholipid transfer protein Isoenzymes Dextran chemistry Human plasma Recombinant DNA Carrier Proteins |
| Zdroj: | Journal of Lipid Research, Vol 45, Iss 2, Pp 387-395 (2004) |
| ISSN: | 0022-2275 |
| DOI: | 10.1194/jlr.d300023-jlr200 |
| Popis: | Human plasma contains two forms of phospholipid transfer protein (PLTP), one catalytically active [high-activity PLTP (HA-PLTP)] and the other a low-activity (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 pretreatment 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 immunoaffinity chromatography, and dextran sulfate-CaCl2 precipitation. All four methods demonstrated that approximately 60% of plasma PLTP represents LA-PLTP and 40% represents 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 +/- SD) (range, 2.78-10.06 mg/l) and the mean activity was 5.84 +/- 1.39 micromol/ml/h (range, 3.21-11.15 micromol/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, whereas that of LA-PLTP (mean, 3.94 +/- 1.4 mg/l) showed no correlation with phospholipid transfer activity. |
| Databáze: | OpenAIRE |
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