Solid-phase enzyme immunoassay of urokinase using monoclonal antibodies

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.

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Neopterin measured in serum and tissue culture supernates by a competitive enzyme-linked immunosorbant assay.

Clinical Chemistry ◽

10.1093/clinchem/35.7.1467 ◽

1989 ◽

Vol 35 (7) ◽

pp. 1467-1471 ◽

Cited By ~ 10

Author(s):

M Barak ◽

D Merzbach ◽

N Gruener

Keyword(s):

Tissue Culture ◽

Correlation Coefficient ◽

Cell Cultures ◽

Solid Phase ◽

Activated Macrophages ◽

Immunosorbant Assay ◽

Analytical Recovery ◽

Enzyme Linked Immunosorbant Assay

Abstract In this solid-phase competitive enzyme-linked immunosorbant assay for neopterin (a product of activated macrophages) in serum or supernates of cell cultures, we incubate antiserum to neopterin with standards or samples in the presence of solid-phase-bound conjugate of carrier-neopterin. Incubation with second antibody labeled with peroxidase then follows, before reaction with substrate. Analytical recovery, precision, and sensitivity of this method are similar to those of other immunoassays. The assay range is between 0.4 and 100 micrograms of neopterin per liter. Results obtained by this method compared with those from a conventional radioimmunoassay gave a correlation coefficient of 0.974.

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Rapid immunotyping of Chlamydia trachomatis with monoclonal antibodies in a solid-phase enzyme immunoassay.

Journal of Clinical Microbiology ◽

10.1128/jcm.22.4.609-613.1985 ◽

1985 ◽

Vol 22 (4) ◽

pp. 609-613 ◽

Cited By ~ 18

Author(s):

R C Barnes ◽

S P Wang ◽

C C Kuo ◽

W E Stamm

Keyword(s):

Monoclonal Antibodies ◽

Chlamydia Trachomatis ◽

Enzyme Immunoassay ◽

Solid Phase

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Generation of Monoclonal Antibodies to Prostatic Acid Phosphatase Isoenzyme 2 And Application in Solid-Phase Enzyme Immunoassay

Biotechnology and Applied Biochemistry ◽

10.1111/j.1470-8744.1989.tb00052.x ◽

1989 ◽

Vol 11 (1) ◽

pp. 89-95 ◽

Cited By ~ 3

Author(s):

CY Kuo ◽

KW Chen ◽

J. Fu ◽

KW Lam ◽

CY Lee

Keyword(s):

Monoclonal Antibodies ◽

Acid Phosphatase ◽

Enzyme Immunoassay ◽

Solid Phase ◽

Prostatic Acid Phosphatase

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Microimmunoassay permits determination of concentrations in immunohistochemistry controls.

Journal of Histochemistry & Cytochemistry ◽

10.1177/34.4.2419398 ◽

1986 ◽

Vol 34 (4) ◽

pp. 543-545

Author(s):

D J Goldstein ◽

M M Davis ◽

R K Naviaux ◽

T L Dailey ◽

T M Ulbright

Keyword(s):

Monoclonal Antibodies ◽

Immunosorbant Assay ◽

Enzyme Linked Immunosorbant Assay

A competition enzyme-linked immunosorbant assay (CELIA) can be used to determine the amount of antigen needed to immunoabsorb an antiserum. This is demonstrated using both polyclonal and monoclonal antibodies against human amylase. This method is of especial value when the tissue is particularly difficult to obtain, and especially valuable when CELIA has already been completed.

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Production of non-overlapped monoclonal antibodies against hepatitis B surface antigen and their use in two-site enzyme linked immunosorbant assay (ELISA)

Archives of Pharmacal Research ◽

10.1007/bf02857759 ◽

1988 ◽

Vol 11 (4) ◽

pp. 261-265 ◽

Cited By ~ 2

Author(s):

Jeung Ah Jang ◽

Won Bae Kim ◽

Won Bong Park ◽

Eung Chil Choi ◽

Byong Kak Kim

Keyword(s):

Monoclonal Antibodies ◽

Hepatitis B ◽

Surface Antigen ◽

Hepatitis B Surface Antigen ◽

Immunosorbant Assay ◽

Enzyme Linked Immunosorbant Assay

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A solid-phase enzyme immunoassay for the detection of tetanus toxin using human and murine monoclonal antibodies

Journal of Basic Microbiology ◽

10.1002/jobm.3620310212 ◽

1991 ◽

Vol 31 (2) ◽

pp. 135-140 ◽

Cited By ~ 3

Author(s):

Stephan T. Kiessig ◽

Christian Hentschel ◽

Sigbert Jahn ◽

Michael Mehl ◽

Roland Starke ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Enzyme Immunoassay ◽

Tetanus Toxin ◽

Solid Phase ◽

Murine Monoclonal Antibodies

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Effects of deglycosylation of human thyroperoxidase on its enzymatic activity and immunoreactivity

Journal of Endocrinology ◽

10.1677/joe.0.1320317 ◽

1992 ◽

Vol 132 (2) ◽

pp. 317-323 ◽

Cited By ~ 23

Author(s):

A. Giraud ◽

J.-L. Franc ◽

Y. Long ◽

J. Ruf

Keyword(s):

Monoclonal Antibodies ◽

Enzymatic Activity ◽

Tertiary Structure ◽

Polyclonal Antibodies ◽

Thyroid Peroxidase ◽

Immunosorbant Assay ◽

Autoimmune Thyroid ◽

Pngase F ◽

Enzyme Linked Immunosorbant Assay ◽

Microsomal Antigen

ABSTRACT Thyroid peroxidase (TPO) is a glycoprotein enzyme which catalyses the iodination of thyroglobulin and the coupling of iodinated tyrosines. Human TPO (hTPO) is the microsomal antigen recognized by the autoantibodies in the serum of patients with autoimmune thyroid disease. An active detergent-solubilized immunoaffinitypurified hTPO was deglycosylated, either by peptide N-glycosidase F (PNGase F) or by endo-β-N-acetylglucosaminidase H (endo H), and the enzymatic activity and immunoreactivity of the native and degylcosylated forms were compared. Electrophoretic controls and affinoblotting with concanavalin A showed that deglycosylation was not total and that it was more pronounced with endo H than with PNGase F. The enzymatic activity of hTPO was inhibited by endo H deglycosylation, but not by PNGase F deglycosylation; this inhibition was not due to aggregation and/or insolubilization of the molecule subsequent to deglycosylation. Immunoreactivity was monitored by enzyme-linked immunosorbant assay (ELISA) with 13 mouse monoclonal antibodies, rabbit polyclonal antibodies and antibodies from serum of patients with Hashimoto's thyroiditis. In contrast with enzymatic activity, immunoreactivity was not modified or was slightly enhanced (with four monoclonal antibodies) by deglycosylation. The results indicate that strong, if not total, deglycosylation induces a modification of the tertiary structure of hTPO, which affects the enzymatic site but does not modify markedly the epitopes implicated in the recognition of the molecule by the antibodies tested. Journal of Endocrinology (1992) 132, 317-323

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Potential of monoclonal antibodies to improve therapeutic monitoring of cyclosporine.

Clinical Chemistry ◽

10.1093/clinchem/33.1.32 ◽

1987 ◽

Vol 33 (1) ◽

pp. 32-37 ◽

Cited By ~ 40

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.

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