A sandwich enzyme immunoassay for the determination of neutrophil lipocalin in body fluids.
Human neutrophil lipocalin was purified from human buffycoat. A polyclonal antibody was obtained by immunisation of rabbits. The antibody reacted with the free lipocalin as well as with the PMNL-gelatinase bound protein. This antibody was used to establish a sensitive sandwich-ELISA for the determination of the protein in body fluids using the biotin/streptavidin system. The mean intra-assay C.V. was 2.3% and the mean inter-assay C.V. 6.7%. The recovery in human plasma was determined to be 98.8%. The ELISA allowed the determination of the protein in the concentration range 0.2-25 micrograms/l. Measurement of the neutrophil lipocalin concentration showed that human plasma of healthy donors contained 9.7 +/- 81 micrograms/l (n = 122) and that the concentrations in serum were significantly higher (P < 0.001) with 133 +/- 90 micrograms/l (n = 122). Neutrophil lipocalin was also found in the urine of healthy donors (8.1 micrograms/l; n = 9). Very high concentrations of this lipocalin were found in the synovial fluids of patients suffering from inflammatory rheumatoid arthritis (1.7 +/- 1.4 mg/l; n = 37).
Bläser
J
J
Faculty of Chemistry, Department of Biochemistry, University of Bielefeld, Germany.
Triebel
S
S
Tschesche
H
H
D000209:Acute-Phase Proteins; D000328:Adult; D000906:Antibodies / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000378:metabolism; D002352:Carrier Proteins / Q000097:blood* / Q000276:immunology / Q000652:urine; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D000071068:Lipocalin-2; D054834:Lipocalins; D008297:Male; D009504:Neutrophils / Q000737:chemistry*; D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins; D013582:Synovial Fluid / Q000737:chemistry*
D000209:Acute-Phase Proteins; D000906:Antibodies; D002352:Carrier Proteins; C489506:LCN2 protein, human; D000071068:Lipocalin-2; D054834:Lipocalins; D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins