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

  • Author(s): Miyashita, Kazuya
  • Kobayashi, Junji
  • Imamura, Shigeyuki
  • Kinoshita, Noriaki
  • Stanhope, Kimber L
  • Havel, Peter J
  • Nakajima, Katsuyuki
  • Machida, Tetsuo
  • Sumino, Hiroyuki
  • Nara, Makoto
  • Murakami, Masami
  • et al.
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.

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