Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins

A sensitive and specific double antibody radio-immunoassay for the major apolipoprotein (apoB) of rhesus (Macaca mulatta) serum very low density lipoprotein (VLDL) and low density lipoprotein (LDL) is described. The anti-serum was raised to LDL (d 1.030–1.040 g/ml) and the LDL2 (d 1.020–1.050 g/ml)...

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Main Authors: J B Karlin, D J Juhn, G Fless, A M Scanu, A H Rubenstein, MD
Format: Article
Language:English
Published: Elsevier 1978-02-01
Series:Journal of Lipid Research
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S0022227520415585
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author J B Karlin
D J Juhn
G Fless
A M Scanu
A H Rubenstein, MD
author_facet J B Karlin
D J Juhn
G Fless
A M Scanu
A H Rubenstein, MD
author_sort J B Karlin
collection DOAJ
description A sensitive and specific double antibody radio-immunoassay for the major apolipoprotein (apoB) of rhesus (Macaca mulatta) serum very low density lipoprotein (VLDL) and low density lipoprotein (LDL) is described. The anti-serum was raised to LDL (d 1.030–1.040 g/ml) and the LDL2 (d 1.020–1.050 g/ml) was labeled with 125I by the chloramine-T or iodine monochloride method. The assay, which was sensitive to 0.02–0.5 µg of LDL2, had an inter-assay coefficient of variation of 4.5%. This assay was successfully used to measure apoB in the whole serum and low density lipoproteins of control monkeys maintained on a standard Purina monkey chow (PMC) diet and of three groups of monkeys fed atherogenic diets: an “average American diet,” a 25% peanut oil and 2% cholesterol-supplemented PMC diet, and a 25% coconut oil and 2% cholesterol-supplemented PMC diet. The control monkeys (n = 13) had a serum cholesterol of 146 ± 28 mg/dl and an apoB of 50 ± 18 mg/dl. In the monkeys maintained on the atherogenic diets the serum apoB was elevated: 103 ± 28 mg/dl (American), 102 ± 35 mg/dl (peanut oil), and 312 ± 88 mg/dl (coconut oil). The values for serum total cholesterol were 333 ± 65 mg/dl (American), 606 ± 212 mg/dl (peanut oil), and 864 ± 233 mg/dl (coconut oil) and were elevated relative to controls (P < 0.001). For each of the diets, total serum cholesterol correlated with serum apoB (P < 0.001). The slopes of the regression lines of serum apoB vs. cholesterol for the monkeys on the PMC, American, and coconut oil diets were similar (m = 0.531, 0.401, and 0.359, respectively), but differed from that of monkeys on the peanut oil diet (m = 0.121). The immunoreactivities of rhesus and human LDL were compared using specific antisera raised against these antigens. In homologous assay systems, monkey and human LDL exhibited unique immunological determinants. The same results were obtained with the delipidated preparations of the two LDLs using antisera raised against either monkey or human apoB. Crossover studies using a heterologous tracer with each anti-serum resulted in the selection of a specific population of antibodies directed against antigenic sites shared by these two LDL species.
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spelling doaj.art-4141fa7f1d2a4e43ba45c1eb80116b602022-12-21T21:49:45ZengElsevierJournal of Lipid Research0022-22751978-02-01192197206Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteinsJ B Karlin0D J Juhn1G Fless2A M Scanu3A H Rubenstein, MD4Department of Medicine, The University of Chicago, Chicago, IL 60637Department of Medicine, The University of Chicago, Chicago, IL 60637Department of Medicine, The University of Chicago, Chicago, IL 60637Department of Medicine, The University of Chicago, Chicago, IL 60637Department of Medicine, The University of Chicago, Chicago, IL 60637; Department of Medicine, The University of Chicago, Box 435, 950 East 59th St., Chicago, IL 60637; Reprint requests should be addressed to Arthur H. Rubenstein, M. D.A sensitive and specific double antibody radio-immunoassay for the major apolipoprotein (apoB) of rhesus (Macaca mulatta) serum very low density lipoprotein (VLDL) and low density lipoprotein (LDL) is described. The anti-serum was raised to LDL (d 1.030–1.040 g/ml) and the LDL2 (d 1.020–1.050 g/ml) was labeled with 125I by the chloramine-T or iodine monochloride method. The assay, which was sensitive to 0.02–0.5 µg of LDL2, had an inter-assay coefficient of variation of 4.5%. This assay was successfully used to measure apoB in the whole serum and low density lipoproteins of control monkeys maintained on a standard Purina monkey chow (PMC) diet and of three groups of monkeys fed atherogenic diets: an “average American diet,” a 25% peanut oil and 2% cholesterol-supplemented PMC diet, and a 25% coconut oil and 2% cholesterol-supplemented PMC diet. The control monkeys (n = 13) had a serum cholesterol of 146 ± 28 mg/dl and an apoB of 50 ± 18 mg/dl. In the monkeys maintained on the atherogenic diets the serum apoB was elevated: 103 ± 28 mg/dl (American), 102 ± 35 mg/dl (peanut oil), and 312 ± 88 mg/dl (coconut oil). The values for serum total cholesterol were 333 ± 65 mg/dl (American), 606 ± 212 mg/dl (peanut oil), and 864 ± 233 mg/dl (coconut oil) and were elevated relative to controls (P < 0.001). For each of the diets, total serum cholesterol correlated with serum apoB (P < 0.001). The slopes of the regression lines of serum apoB vs. cholesterol for the monkeys on the PMC, American, and coconut oil diets were similar (m = 0.531, 0.401, and 0.359, respectively), but differed from that of monkeys on the peanut oil diet (m = 0.121). The immunoreactivities of rhesus and human LDL were compared using specific antisera raised against these antigens. In homologous assay systems, monkey and human LDL exhibited unique immunological determinants. The same results were obtained with the delipidated preparations of the two LDLs using antisera raised against either monkey or human apoB. Crossover studies using a heterologous tracer with each anti-serum resulted in the selection of a specific population of antibodies directed against antigenic sites shared by these two LDL species.http://www.sciencedirect.com/science/article/pii/S0022227520415585low density lipoprotein radioimmunoassayatherosclerosishypercholesterolemia
spellingShingle J B Karlin
D J Juhn
G Fless
A M Scanu
A H Rubenstein, MD
Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins
Journal of Lipid Research
low density lipoprotein radioimmunoassay
atherosclerosis
hypercholesterolemia
title Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins
title_full Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins
title_fullStr Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins
title_full_unstemmed Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins
title_short Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins
title_sort measurment of rhesus monkey macaca mulatta apolipoprotein b in serum by radioimmunoassay comparison of immunoreactivities of rhesus and human low density lipoproteins
topic low density lipoprotein radioimmunoassay
atherosclerosis
hypercholesterolemia
url http://www.sciencedirect.com/science/article/pii/S0022227520415585
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