Radioimmunoassay of beta lipoprotein—protein of rat serum

Eaton, R. Philip; Kipnis, David

doi:10.1172/jci106104

Cited by 28 publications

(10 citation statements)

References 24 publications

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“…Tissue contents of ApoA-I (28) and ApoB (31,37) were assessed by radioimmunoassay. Mucosal scrapings ofproximal jejunum were homogenized in three volumes of 0.05 M barbital pH 8.6, 1% Triton X-100 and centrifuged at 105,000 g for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Intestinal apoproteins during fat absorption.

Schonfeld¹,

Bell²,

Alpers³

1978

J. Clin. Invest.

129

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Section: Methodsmentioning

confidence: 99%

Intestinal apoproteins during fat absorption.

Schonfeld¹,

Bell²,

Alpers³

1978

J. Clin. Invest.

129

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“…J. ); all dilutions were made in 3% BSA in barbital buffer (17). The the tubes were centrifuged, the precipitates washed with cold barbital buffer and counted in a Nuclear-Chicago gamma spectrometer (Nuclear-Chicago Corp., Des Plaines, Ill.).…”

Section: Methodsmentioning

confidence: 99%

“…lipid than of their protein moieties, because, for most of the apolipoproteins, the available methodology is cumbersome, time-consuming, and applicable only to a relatively small number of samples at a time (10)(11)(12)(13)(14)(15). Recently, simpler assays capable of measuring apolipoprotein B (ApoB)1 in human low density lipoproteins (LDL) (16,17) and in rat plasma have become available (18). However, the assay used for assessing human ApoB depends upon the diffusion lAbbreviations used in this paper: ApoB, apolipoprotein B; BSA, bovine serum albumin; d-LDL, delipidated low density lipoprotein; LDL, low density lipoprotein; RIA, radioimmunoassay; SDS, sodium dodecyl sulfate; VLDL, very low density lipoprotein.…”

Section: Introductionmentioning

confidence: 99%

Assay of Total Plasma Apolipoprotein B Concentration in Human Subjects

Schonfeld¹,

Lees²,

George³

et al. 1974

J. Clin. Invest.

171

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A B S T R A C T We have developed a double antibody radioimmunoassay (RIA) for human apolipoprotein B (ApoB). The assay measures not only the ApoB content of 8-lipoproteins (low density lipoproteins [LDL]) but also that contained in the other lipoproteins in plasma.Purified lymph and plasma chylomicrons and plasma very low density lipoproteins (VLDL) produced displacement curves in the assay system which paralleled those produced by pure LDL. Thus, the ApoB found in chylomicrons, VLDL, and LDL were immunologically identical. ApoB accounted for about 25 and 35%, respectively, of the total protein of chylomicrons and VLDL by RIA. VLDL and LDL preparations from normal and hyperlipoproteinemic subjects also produced parallel displacement curves, suggesting that the ApoB of normal and hyperlipoproteinemic subjects were immunologically identical. High density lipoproteins and abetalipoproteinemic plasma displaced no counts, nor did the sera of several animal species produce any useful displacement curves in this system.The fasting total plasma ApoB concentration of normal subjects was 83±16 mg/dl (mean+SD). ApoB levels were high in Type II (162±16), and less so in Type IV (112±24) and Type V (105±17). When plasma ApoB concentration in Type IV patients was graphed against plasma glycerides, two subpopulations, which may represent different genetic or biochemical subgroups, were apparent. ApoB concentration in individuals on constant diet and drug regimen was stable over weeks to months. Greater than 90% of ApoB of normal and Type II subjects was in the d> 1.006 plasma fraction. By contrast, only 50-80% of ApoB was in the d > 1.006 fraction in Types IV and V. Thus, hypertriglyceridemia was associated primarily with a redistribution of ApoB to the lighter density fractions; by contrast, in hypercholesterolemia absolute ApoB concentration was markedly increased.

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“…Apo B of LDL was measured in a single radial immunodiffusion assay [6]. Later Eaton & Kipnis [7] developed a radioimmunoassay for the quantification of apo B in rat serm. While the present work was in progress, Gotto et al [8] , Schonfeld et al [9] , Albers et al…”

Section: Introductionmentioning

confidence: 99%

Measurement of serum apolipoprotein B by radioimmunoassay

Karlin

Juhn

Scanu

et al. 1978

Eur J Clin Investigation

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A sensitive and specific double antibody radioimmunoassay for the major apolipoprotein (apo B) of human serum very low density lipoprotein (VLDL) and low density lipoprotein (LDL) is described. Using anti-LDL and anti-apo B antibodies the immunoreactivity of LDL and apo B were compared. Human LDL and its isolated apo B were not immunologically identical when each antiserum was used with its homologous label; a population of antibodies was selected which reacted with antigenic sites unique to the antigen itself as well as to those which were common to the closely related protein. When the heterologous label was used with either antiserum, a population of antibodies directed against antigenic sites shared by the LDL and apo B molecules was selected.Apo B in sera samples can be measured using either anti-LDL or anti-apo B antibodies provided that intact LDL was used for preparation of the iodinated tracer and standard. Serum apo B levels in healthy normolipidaemic males and females were 0.93 f 0.25 g/l (range 0.58-1.39) and 0.90 f 0.15 g/l (range 0.58-1.12), respectively. The total cholesterol and apo B, and phospholipid and apo B concentrations for both males and females were significantly correlated (P < 0.05). In another normolipidaemic population (n = 52), total serum apo B values correlated positively with LDL cholesterol (r = 0.92, P < 0.001).Apo B was measured in sera from patients with abetalipoproteinaemia, familial hypercholesterolaemia and Tangiers disease. Apo B was not detected in the serum of subjects with abetalipoproteinaemia, while the apo B level in the familial hypercholesterolaemic subjects was significantly elevated (range 3.26-4.94 g/l) compared to normals (P

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Radioimmunoassay of beta lipoprotein—protein of rat serum

Cited by 28 publications

References 24 publications

Intestinal apoproteins during fat absorption.

Intestinal apoproteins during fat absorption.

Assay of Total Plasma Apolipoprotein B Concentration in Human Subjects

Measurement of serum apolipoprotein B by radioimmunoassay

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