Measurement of human plasma phospholipid transfer protein by sandwich ELISA

Clin Chem. 2000 Sep;46(9):1357-64.

Abstract

Background: Plasma phospholipid transfer protein (PLTP) plays a central role in the remodeling of HDLs. Reliable and accurate methods for assaying PLTP concentration are required.

Methods: A sandwich ELISA for PLTP has been developed, using two monoclonal antibodies against recombinant human PLTP (rhPLTP) expressed in Chinese hamster ovary cells. The ELISA allows for the quantification of PLTP in the range 0.625-15.0 ng/assay (1.2-30.0 mg/L). Intra- and interassay CVs were <3.0% and <4.2% respectively. The assay was used to quantify plasma PLTP concentrations in 132 Japanese subjects (75 males and 57 females).

Results: PLTP concentrations were 12.0 +/- 3. 0 mg/L (mean +/- SD; range, 4.9-20.5 mg/L). No sex difference was observed. Plasma PLTP concentration was positively correlated with HDL-cholesterol (r = 0.72; P: <0.001), apolipoprotein (apo) A-I (r = 0.62; P: <0.001) and HDL(2)-cholesterol (r = 0.72; P: <0.001), and was negatively correlated with triacylglycerol (r = -0.45; P: <0. 001). There was no correlation with plasma apo A-II. These results agree with other evidence that plasma PLTP is associated with large apo A-I-containing lipoproteins. There was no correlation (r = -0. 01) between plasma PLTP and plasma phosphatidylcholine transfer activity (range, 3.5-10.5 micromol. mL(-1). h(-1)), suggesting that PLTP may exist in active and inactive forms.

Conclusion: This new ELISA will be of value for further studies of PLTP in health and disease.

MeSH terms

  • Adult
  • Animals
  • Antibodies, Monoclonal
  • CHO Cells
  • Calibration
  • Carrier Proteins / blood*
  • Carrier Proteins / immunology
  • Carrier Proteins / metabolism
  • Cricetinae
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Humans
  • Male
  • Membrane Proteins / blood*
  • Membrane Proteins / immunology
  • Membrane Proteins / metabolism
  • Middle Aged
  • Phospholipid Transfer Proteins*
  • Phospholipids / metabolism*
  • Recombinant Proteins / immunology

Substances

  • Antibodies, Monoclonal
  • Carrier Proteins
  • Membrane Proteins
  • Phospholipid Transfer Proteins
  • Phospholipids
  • Recombinant Proteins