A monoclonal antibody-based enzyme immunoassay for the determination of glycated high density lipoproteins

Abstract I describe a method for measuring high-density lipoprotein phospholipids. Magnesium chloride and dextran sulfate are used to precipitate all low-density and very-low-density lipoproteins. The supernate contains only high-density lipoproteins, the phospholipid concentration of which is determined by an enzymic method. The precision of the method (CV) is 2.35% (10 repeated assays), and the mean value for HDL-phospholipids was 1006 (SD 248) mg/L for 30 apparently healthy subjects. I used electrophoresis and enzymic color development to confirm the presence of HDL-phospholipids. Results are compared with those obtained by an ultracentrifugation method.

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Determination of Domoic Acid in Japanese Mussels by Enzyme Immunoassay

Journal of AOAC International ◽

10.1093/jaoac/83.6.1384 ◽

2000 ◽

Vol 83 (6) ◽

pp. 1384-1386 ◽

Cited By ~ 11

Author(s):

Kentaro Kawatsu ◽

Yonekazu Hamano ◽

Tamao Noguchi

Keyword(s):

Monoclonal Antibody ◽

Enzyme Immunoassay ◽

Domoic Acid ◽

Immunoaffinity Chromatography ◽

Shellfish Poisoning ◽

Mussel Tissue ◽

Blue Mussels ◽

Amnesic Shellfish Poisoning ◽

Low Concentrations

Abstract Ten samples of commercial blue mussels (Mytilus edulis) from Japan were analyzed for domoic acid by an indirect competitive enzyme immunoassay (idc–EIA) based on an anti-domoic acid monoclonal antibody. Domoic acid was found in all samples at low concentrations (0.11–1.81 ng/g mussel tissue). The presence of domoic acid was confirmed by liquid chromatography coupled with immunoaffinity chromatography using an anti-domoic acid monoclonal antibody as ligand. To our knowledge, this is the first reported detection of domoic acid, a causative agent of amnesic shellfish poisoning, in Japanese mussels.

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Densitometry of phosphatidylcholine and sphingomyelin in high-density lipoproteins.

Clinical Chemistry ◽

10.1093/clinchem/29.7.1435 ◽

1983 ◽

Vol 29 (7) ◽

pp. 1435-1437 ◽

Cited By ~ 4

Author(s):

G Schmitz ◽

H U Jabs ◽

G Assmann

Keyword(s):

Sulfuric Acid ◽

Phosphotungstic Acid ◽

High Density ◽

High Density Lipoproteins ◽

Low Concentrations ◽

Layer Chromatography ◽

Inexpensive Technique ◽

Native Plasma ◽

Enzymatic Methods

Abstract We describe the quantitative densitometric determination of phosphatidylcholine (PC) and sphingomyelin (SP) in human serum after precipitation with phosphotungstic acid/MgCl2 and use of thin-layer chromatography. After development, chromatographic plates were charred with methanolic sulfuric acid and MnCl2 and scanned by direct reflectance densitometry in an automated densitometric system interfaced to a basic programmable computing integrator. The method is sensitive enough to detect abnormally low concentrations of PC and SP in high-density lipoproteins. The accuracy of the method was tested either with the Bartlett phosphorus assay or with enzymatic methods for PC and SP; correlations of the described method with the enzymatic determinations were r = 0.93 and 0.88, respectively. Day-to-day precision (CV) for the phospholipid determination was 8.6% for PC and 12.2% for SP. The major advantage of this inexpensive technique is that native plasma or serum or the serum supernate after precipitation can be used without prior delipidation. With this technique serum high-density lipoproteins had PC values of 1.08 (SD 0.32) mmol/L in men (n = 158) and 1.12 (SD 0.37) mmol/L in women (n = 192); similarly, SP values were 0.23 (SD 0.07) mmol/L in the men and 0.23 (SD 0.08) mmol/L in the women. The differences by sex are not significant.

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Determination of high-density lipoproteins: comparison of HDL-cholesterol, HDL-phospholipids, and HDL-apolipoprotein A-I.

Clinical Chemistry ◽

10.1093/clinchem/27.2.348 ◽

1981 ◽

Vol 27 (2) ◽

pp. 348-348 ◽

Cited By ~ 4

Author(s):

P Weisweiler ◽

B Sperl ◽

P Schwandt

Keyword(s):

Hdl Cholesterol ◽

High Density ◽

High Density Lipoproteins ◽

Apolipoprotein A

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Specific immunoassay of alpha-amylase isoenzymes in human serum.

Clinical Chemistry ◽

10.1093/clinchem/31.8.1331 ◽

1985 ◽

Vol 31 (8) ◽

pp. 1331-1334 ◽

Cited By ~ 23

Author(s):

M Gerber ◽

K Naujoks ◽

H Lenz ◽

W Gerhardt ◽

K Wulff

Keyword(s):

Monoclonal Antibody ◽

Human Serum ◽

Enzyme Immunoassay ◽

Amylase Activity ◽

Alpha Amylase ◽

Pancreatic Amylase ◽

Routine Method ◽

Salivary Amylase ◽

Amylase Isoenzymes

Abstract A monoclonal antibody (66C7) was prepared that specifically binds human salivary amylase (EC 3.2.1.1); it cross reacts with human pancreatic amylase by less than 1%. Two procedures are described for determination of isoamylases in human serum with this antibody: an enzyme immunoassay for determining amylase of salivary origin, and a routine method in which this amylase is immunoprecipitated and the remaining (pancreatic) amylase activity is assayed. Results by the two methods correlate well.

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