Potential of monoclonal antibodies to improve therapeutic monitoring of cyclosporine.

1987 ◽  
Vol 33 (1) ◽  
pp. 32-37 ◽  
Author(s):  
V Quesniaux ◽  
R Tees ◽  
M H Schreier ◽  
G Maurer ◽  
M H van Regenmortel
Keyword(s):  
Alkaline Phosphatase ◽  
Monoclonal Antibodies ◽  
Enzyme Immunoassay ◽  
Cyclosporine A ◽  
Solid Phase ◽  
Therapeutic Monitoring ◽  
High Affinity ◽  
Polyclonal Antisera ◽  
Direct Solid

Abstract We show that monitoring of cyclosporine by immunoassay could be improved by using monoclonal antibodies (MAbs) of restricted specificity instead of polyclonal antisera that recognize both unmodified cyclosporine and its metabolites. MAbs with high affinity for cyclosporine have been prepared and characterized. We tested their ability to discriminate between native cyclosporine and its metabolites in indirect solid-phase enzyme immunoassay with a set of cyclosporine metabolites modified at residues 1, 4, 6, and 9 (corresponding to the six known sites of metabolism of cyclosporine). All the metabolites tested were detected by MAb1 at least 15- to 1000-fold less well than unmodified cyclosporine. A second MAb recognized unmodified cyclosporine and most of its metabolites equally well. Both MAbs retained their activity when coupled to alkaline phosphatase and could therefore be used in a direct solid-phase enzyme immunoassay.

Use of monoclonal antibodies to detect human placental alkaline phosphatase.

1983 ◽  
Vol 29 (1) ◽  
pp. 115-119 ◽  
Author(s):  
G De Groote ◽  
P De Waele ◽  
A Van de Voorde ◽  
M De Broe ◽  
W Fiers
Keyword(s):  
Monoclonal Antibody ◽  
Alkaline Phosphatase ◽  
Monoclonal Antibodies ◽  
Solid Phase ◽  
Enzymatic Reactions ◽  
Placental Alkaline Phosphatase ◽  
Human Placental Alkaline Phosphatase ◽  
Human Sera ◽  
Starch Gel ◽  
Alkaline Phosphatase Isoenzyme

Abstract Convenient, sensitive, and specific solid-phase immunoassays involving monoclonal antibody are described for the determination of human placental alkaline phosphatase (hPLAP). An endogenous enzyme immunoassay combined the specificity of the immunological and the enzymatic reactions. Alternatively, a solid-phase "sandwich" radioimmunoassay involving immobilized polyclonal rabbit anti-hPLAP in combination with iodinated monoclonal antibody provided some additional advantages. Both tests can be used to detect hPLAP from various sources, e.g., in human sera during pregnancy or as a tumor marker. The radioimmunoassay detected an increase in hPLAP at nine weeks of gestation. We discuss the use of monoclonal antibodies for the differentiation of different alkaline phosphatase isoenzyme types by electrophoresis on starch gel.

scholarly journals High-affinity monoclonal antibodies for aflatoxins and their application to solid-phase immunoassays.

1984 ◽  
Vol 81 (24) ◽  
pp. 7728-7731 ◽  
Author(s):  
J. D. Groopman ◽  
L. J. Trudel ◽  
P. R. Donahue ◽  
A. Marshak-Rothstein ◽  
G. N. Wogan
Keyword(s):  
Monoclonal Antibodies ◽  
Solid Phase ◽  
High Affinity

Solid-phase enzyme immunoassay of urokinase using monoclonal antibodies

1983 ◽  
Vol 3 (4) ◽  
pp. 381-388 ◽  
Author(s):  
P. Hérion ◽  
D. Portetelle ◽  
J. -D. Franssen ◽  
J. Urbain ◽  
A. Bollen
Keyword(s):  
Monoclonal Antibodies ◽  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Biological Fluids ◽  
Immunosorbant Assay ◽  
Rapid Procedure ◽  
Preliminary Purification ◽  
Enzyme Linked Immunosorbant Assay

Using two monoclonal antibodies directed against urokinase, we have developed a micro enzyme-linked immunosorbant assay (ELISA) to detect and measure urokinase in biological fluids. The system presents the following characteristics: simple and rapid procedure, reproducibility, sensitivity (urokinase levels down to 1 ng/ml) and evaluation of the enzyme in biological fluids such as urine, pleural elfusions, and ascitic fluids without preliminary purification.

Screening of monoclonal antibodies for hCG for affinity and specificity

1988 ◽  
Vol 66 (8) ◽  
pp. 910-916
Author(s):  
Lorraine Leblond ◽  
Édith Lemieux ◽  
Praful Patel ◽  
Philippe Crine
Keyword(s):  
Monoclonal Antibodies ◽  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
Solid Phase ◽  
Immunoradiometric Assay ◽  
High Affinity ◽  
Heterologous Antigens

Using human chorionic gonadotropin (hCG) as a model polypeptide, we have developed a strategy that allows the direct screening of supernatant fluids from hybridomas for the presence of monoclonal antibodies of high affinity and predefined specificities. The assay evaluates the competition between 125I-labeled and unlabeled homologous or heterologous antigens in a solid-phase two-site immunoradiometric assay. This assay is fast and accurate, and is of general use provided the antigen of interest can be purified in nanomolar quantities. This strategy led to the isolation of nine new monoclonal antibodies for hCG, two of which could be used for elaborating a sensitive two-site immunoradiometric assay for this hormone.

scholarly journals Solid-phase enzyme immunoassay for immunoglobulin M antibodies to cytomegalovirus

1977 ◽  
Vol 5 (6) ◽  
pp. 629-634
Author(s):  
H Schmitz ◽  
H W Doerr ◽  
D Kampa ◽  
A Vogt
Keyword(s):  
Alkaline Phosphatase ◽  
Horseradish Peroxidase ◽  
Enzyme Immunoassay ◽  
Indirect Immunofluorescence ◽  
Solid Phase ◽  
Immunoglobulin M ◽  
Nuclear Antigen ◽  
Immunofluorescence Method ◽  
Infected Cells

A sensitive, solid-phase enzyme immunoassay for the detection of immunoglobulin M antibodies to cytomegalovirus is described. The results of the enzyme immunoassay correlated well with those obtained by an indirect immunofluorescence method. Horseradish peroxidase proved to be a more sensitive label than alkaline phosphatase. Nonspecific reactions, occurring with commercially available cytomegalovirus antigens, could be avoided by using a nuclear antigen prepared from sonically disrupted nuclei of cytomegalovirus-infected cells.

Enzyme immunoassay of antibodies to Sjögren's syndrome B antigen.

1981 ◽  
Vol 27 (8) ◽  
pp. 1341-1345 ◽  
Author(s):  
A M Teppo
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Immunoassay ◽  
Sjögren’S Syndrome ◽  
Sjögren's Syndrome ◽  
Solid Phase ◽  
Sjogren’S Syndrome ◽  
Sjogren's Syndrome ◽  
Nitrophenyl Phosphate ◽  
Purified Antigen ◽  
Sjögren‘S Syndrome

Abstract I describe a solid-phase enzyme immunoassay of IgG-IgA-, and IgM-type antibodies to Sjögren's syndrome B antigen. Polystyrene tubes are coated with the purified antigen. The antibodies are allowed to bind with their antigens, and then detected with alkaline-phosphatase-conjugated anti-human IgG, IgA, or IgM sera. The amount of alkaline phosphatase fixed to the tubes is determined in pH 10.0 diethanolamine buffer at 37 degrees C with p-nitrophenyl phosphate as substrate. The absorbance of the p-nitrophenolate ion liberated in 1 h at 37 degrees C is measured at 406 nm. When rheumatoid factor is present, the values obtained for IgG antibodies are too low, and those for IgM antibodies are too high.

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