A new enzyme-linked immunosorbent assay system for human hepatic triglyceride lipase

Clin Chim Acta. 2013 Sep 23:424:201-6. doi: 10.1016/j.cca.2013.06.016. Epub 2013 Jun 27.

Abstract

Background: The objective of this study was to establish a new sandwich based enzyme linked immunosorbent assay (ELISA) for measuring the protein mass of human hepatic triacylglyceride lipase (HTGL).

Method: Two mouse monoclonal antibodies raised against human HTGL were used for the sandwich ELISA. The post-heparin plasma (PHP) samples obtained at a heparin dose of 50 unit/kg from 124 normolipidemic subjects were used for this ELISA.

Results: The dynamic assay range of the developed ELISA for the HTGL was from 0.47 to 30 ng/ml. The CV was <7% in both intra- and inter-assays, and it did not cross-react with lipoprotein lipase or endothelial lipase (EL). The HTGL concentration in PHP showed a strong correlation with HTGL activity [n=121, r=0.778, p<0.001]. There was a weak relation of HTGL concentration against high-density lipoprotein cholesterol (HDL-C) [n=123, r=-0.229, p=0.011] but no relations against total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), small dense LDL, remnant like particles cholesterol (RLP-C) and RLP-TG were confirmed. Interestingly, a weak but positive correlation between HTGL concentration and EL concentration was shown [n=122, p=0.013, r=0.224].

Conclusion: These results indicate that this new sandwich ELISA for measuring HTGL concentration in PHP can be applied in a daily clinical practice.

Keywords: ELISA; Endothelial lipase; Hepatic triglyceride lipase.

MeSH terms

  • Adult
  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / isolation & purification*
  • CHO Cells
  • Cholesterol, HDL / blood
  • Cholesterol, LDL / blood
  • Cricetulus
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Lipase / blood*
  • Liver / enzymology*
  • Male
  • Mice
  • Obesity / blood*
  • Recombinant Proteins / analysis
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Triglycerides / blood

Substances

  • Antibodies, Monoclonal
  • Cholesterol, HDL
  • Cholesterol, LDL
  • LIPC protein, human
  • Recombinant Proteins
  • Triglycerides
  • Lipase