Quantification of lipoprotein lipase in mouse plasma with a sandwich enzyme-linked immunosorbent assay

采用夹心酶联免疫吸附试验法定量小鼠血浆中的脂蛋白脂肪酶

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作者:Takao Kimura,Kazuya Miyashita,Isamu Fukamachi,Kumiko Fukamachi,Kazumi Ogura,Erina Yokoyama,Katsuhiko Tsunekawa,Takumi Nagasawa,Michael Ploug,Ye Yang,Wenxin Song,Stephen G Young,Anne P Beigneux,Katsuyuki Nakajima,Masami Murakami

Abstract

To support in vivo and in vitro studies of intravascular triglyceride metabolism in mice, we created rat monoclonal antibodies (mAbs) against mouse LPL. Two mAbs, mAbs 23A1 and 31A5, were used to develop a sandwich ELISA for mouse LPL. The detection of mouse LPL by the ELISA was linear in concentrations ranging from 0.31 ng/ml to 20 ng/ml. The sensitivity of the ELISA made it possible to quantify LPL in serum and in both pre-heparin and post-heparin plasma samples (including in grossly lipemic samples). LPL mass and activity levels in the post-heparin plasma were lower in Gpihbp1-/- mice than in wild-type mice. In both groups of mice, LPL mass and activity levels were positively correlated. Our mAb-based sandwich ELISA for mouse LPL will be useful for any investigator who uses mouse models to study LPL-mediated intravascular lipolysis.

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