Kazuya Miyashita1, Isamu Fukamachi1, Tetsuo Machida2, Kiyomi Nakajima2, Stephen G Young3, Masami Murakami2, Anne P Beigneux4, Katsuyuki Nakajima5. 1. Immuno Biological Laboratories, Fujioka, Gunma, Japan. 2. Department of Clinical Laboratory Medicine, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan. 3. Department of Medicine and David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90025, United States; Department of Human Genetics, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90025, United States. 4. Department of Medicine and David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90025, United States. Electronic address: abeigneux@mednet.ucla.edu. 5. Department of Clinical Laboratory Medicine, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan. Electronic address: nakajimak05@ybb.ne.jp.
Abstract
BACKGROUND: Autoantibodies against GPIHBP1, the endothelial cell transporter for lipoprotein lipase (LPL), cause severe hypertriglyceridemia ("GPIHBP1 autoantibody syndrome"). Affected patients have low serum GPIHBP1 and LPL levels. We report the development of a sensitive and specific ELISA, suitable for routine clinical use, to detect GPIHBP1 autoantibodies in serum and plasma. METHODS: Serum and plasma samples were added to wells of an ELISA plate that had been coated with recombinant human GPIHBP1. GPIHBP1 autoantibodies bound to GPIHBP1 were detected with an HRP-labeled antibody against human immunoglobulin. Sensitivity, specificity, and reproducibility of the ELISA was evaluated with plasma or serum samples from patients with the GPIHBP1 autoantibody syndrome. RESULTS: A solid-phase ELISA to detect and quantify GPIHBP1 autoantibodies in human plasma and serum was developed. Spiking recombinant human GPIHBP1 into the samples reduced the ability of the ELISA to detect GPIHBP1 autoantibodies. The ELISA is reproducible and sensitive; it can detect GPIHBP1 autoantibodies in samples diluted by >1000-fold. CONCLUSION: We have developed a sensitive and specific ELISA for detecting GPIHBP1 autoantibodies in human serum and plasma; this assay will make it possible to rapidly diagnose the GPIHBP1 autoantibody syndrome.
BACKGROUND: Autoantibodies against GPIHBP1, the endothelial cell transporter for lipoprotein lipase (LPL), cause severe hypertriglyceridemia ("GPIHBP1 autoantibody syndrome"). Affected patients have low serum GPIHBP1 and LPL levels. We report the development of a sensitive and specific ELISA, suitable for routine clinical use, to detect GPIHBP1 autoantibodies in serum and plasma. METHODS: Serum and plasma samples were added to wells of an ELISA plate that had been coated with recombinant humanGPIHBP1. GPIHBP1 autoantibodies bound to GPIHBP1 were detected with an HRP-labeled antibody against human immunoglobulin. Sensitivity, specificity, and reproducibility of the ELISA was evaluated with plasma or serum samples from patients with the GPIHBP1 autoantibody syndrome. RESULTS: A solid-phase ELISA to detect and quantify GPIHBP1 autoantibodies in human plasma and serum was developed. Spiking recombinant humanGPIHBP1 into the samples reduced the ability of the ELISA to detect GPIHBP1 autoantibodies. The ELISA is reproducible and sensitive; it can detect GPIHBP1 autoantibodies in samples diluted by >1000-fold. CONCLUSION: We have developed a sensitive and specific ELISA for detecting GPIHBP1 autoantibodies in human serum and plasma; this assay will make it possible to rapidly diagnose the GPIHBP1 autoantibody syndrome.
Authors: Kazuya Miyashita; Jens Lutz; Lisa C Hudgins; Dana Toib; Ambika P Ashraf; Wenxin Song; Masami Murakami; Katsuyuki Nakajima; Michael Ploug; Loren G Fong; Stephen G Young; Anne P Beigneux Journal: J Lipid Res Date: 2020-09-18 Impact factor: 5.922
Authors: Stephen G Young; Loren G Fong; Anne P Beigneux; Christopher M Allan; Cuiwen He; Haibo Jiang; Katsuyuki Nakajima; Muthuraman Meiyappan; Gabriel Birrane; Michael Ploug Journal: Cell Metab Date: 2019-07-02 Impact factor: 27.287
Authors: Jens Lutz; Malgorzata Dunaj-Kazmierowska; Sven Arcan; Ursula Kassner; Kazuya Miyashita; Masami Murakami; Michael Ploug; Loren G Fong; Stephen G Young; Katsuyuki Nakajima; Anne P Beigneux Journal: Ann Intern Med Date: 2020-08-11 Impact factor: 25.391
Authors: Ambika P Ashraf; Kazuya Miyashita; Katsuyuki Nakajima; Masami Murakami; Robert A Hegele; Michael Ploug; Loren G Fong; Stephen G Young; Anne P Beigneux Journal: J Clin Lipidol Date: 2020-01-31 Impact factor: 4.766