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

1991 ◽  
Vol 31 (2) ◽  
pp. 135-140 ◽  
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

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.

The Effect of Human Serum on the Binding Activity of Radiolabelled Monoclonal Antibodies

1987 ◽  
Vol 73 (6) ◽  
pp. 547-554
Author(s):  
Silvia Camagni ◽  
Silvana Canevari ◽  
Marina Ripamonti ◽  
Delia Mezzanzanica ◽  
Rosaria Orlandi ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Human Blood ◽  
Solid Phase ◽  
Binding Capacity ◽  
Binding Activity ◽  
Ovarian Carcinomas ◽  
Radiolabelled Monoclonal Antibodies ◽  
In Vitro Stability ◽  
Murine Monoclonal Antibodies

Three murine monoclonal antibodies (MoAbs), MBrl and MOv2 of IgM isotype and MOv8 of IgG isotype, with restricted reactivity for breast or ovarian carcinomas, were labelled with 125I in the perspective of obtaining specific and stable radioimmunopharmaceutical reagents. The radiolabeled MoAbs were analyzed for their « in vitro » stability in human blood. They were incubated at 37 °C for various lengths of time in human or, as a control, in murine blood and their binding capacity was evaluated by solid-phase RIA and compared with that obtained after incubation with buffer. In human blood, serum and plasma, but not with other components such as erythrocytes, leukocytes, HSA and IgG, the MoAbs revealed a loss of binding reactivity which was marked and constant for the IgM MoAbs, and only occasional for the IgG MoAb. In murine serum the decrease was not so rapid. The same change in the binding capacity was observed when the MoAbs were labelled with 3H or 35S, excluding the involvement of dehalogenating mechanisms. In the perspective of using MoAbs for intracavity therapy the effect of ascitic or pleural fluids on their binding activity was also evaluated. The inhibition of the binding reactivity was not as evident and was not related to the MoAb isotype.

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.

A solid-phase enzyme immunoassay for the common and subtypic determinants of hepatitis B surface antigen with monoclonal antibodies

1986 ◽  
Vol 87 (2) ◽  
pp. 203-210 ◽  
Author(s):  
Sadakazu Usuda ◽  
Fumio Tsuda ◽  
Tohru Gotanda ◽  
Katsumi Tachibana ◽  
Motozumi Nomura ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Hepatitis B ◽  
Enzyme Immunoassay ◽  
Surface Antigen ◽  
Solid Phase ◽  
Hepatitis B Surface Antigen ◽  
The Common

scholarly journals Two new murine monoclonal antibodies rised against human IgG

2001 ◽  
Vol 66 (5) ◽  
pp. 323-330 ◽  
Author(s):  
Marijana Petricevic ◽  
Aleksandra Inic ◽  
Ratko Jankov ◽  
Ljiljana Dimitrijevic
Keyword(s):  
Monoclonal Antibodies ◽  
Solid Phase ◽  
Proteolytic Enzymes ◽  
Cross Reactivity ◽  
Dot Blot ◽  
Human Igg ◽  
Western Blots ◽  
Binding Characteristics ◽  
Hybridoma Technology ◽  
Murine Monoclonal Antibodies

Many pathological conditions are accompanied with changes in the concentration of the total IgG or some of its fraction. For this reason there is great interest in the production of reagents specific for IgG. In this paper, the binding characteristics of two new murine monoclonal antibodies (MoAb), assigned MoAb 15 and MoAb 22, are reported. These MoAbs were produced by hybridoma technology. By performing ELISAs and Western blots analyzes, it was demonstrated that both MoAbs interact specifically with human IgG. Cross reactivity with other sera proteins was not observed. In order to precisely localize the epitopes recognized by MoAb 15 and MoAb 22, the Western blots interactions of these MoAbs with electrophoreticaly separated IgG-fragments, obtained by the action of proteolytic enzymes (papain, pepsin, trypsin), were analyzed. According to the results of these experiments, both MoAbs interacted with epitopes in the C 3 domain. The affinity constants, calculated from Scatchard plots of binding ofMoAb15 and MoAb22 to human IgG, wereKa15 = 1.71 106M-1 andKa22 = 2.15 109M-1. According to all these findings,MoAb 15 and MoAb 22 could be used in standard immunochemical techniques. However, the experiments showed that both MoAbs had bad immunoprecipitating properties. In solid phase techniques (ELISAs, Western blot, dot-blot, etc.), their application gave excellent results that highly recommended them for use in these types of analyzes.

Development of enzyme immunoassay for detecting I and II types of shiga-like toxins

2021 ◽  
Vol 29 (5) ◽  
pp. 43-48
Author(s):  
Galina Viktorovna Kuklina ◽  
Denis Valerievich Pechenkin ◽  
Sergei Sergeevich Ipatov ◽  
Andrei Valentinovich Eremkin ◽  
Aleksei Aleksandrovich Kytmanov ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Sandwich Elisa ◽  
Central Research Institute ◽  
Exchange Chromatography ◽  
Hybridoma Cells ◽  
Federal State ◽  
Central Research ◽  
Freeze Dried

Introduction. The aim of the work was development of enzyme immunoassay for detecting I and II types of shiga-like toxins and assessment of it diagnostic properties. Materials and methods. For the research, we used hybridomas producing monoclonal antibodies to shiga-like toxins of types I and II, obtained at the branch of the Federal State Budgetary Institution “48 Central Research Institute” of the Ministry of Defense of Russian Federation (Kirov); BALB/c mice; shiga-like toxins of types I and II. Hybridoma cells were cultured in culture flasks and in the peritoneal cavity of BALB/c mice. Monoclonal antibodies were isolated from ascitic fluids by precipitation with a saturated solution of ammonium sulfate, followed by purification by ion exchange chromatography. The obtained preparations of monoclonal antibodies were used to develop enzyme immunoassay for the detection of shiga-like toxins of types I and II. Specific components of enzyme immunoassay were freeze-dried in a protective environment. Results. As a result of research, preparative quantities of monoclonal antibodies against I and II types of shiga-like toxins were obtained and purified; selection of monoclonal antibodies for sorption on the solid phase and for the synthesis of immunoperoxidase conjugates was carried out. Conclusion. experimental enzyme immunoassay allowing to identify 1 ng/ml I and II types of shiga-like toxins in «sandwich»-ELISA was developed.

Sign in / Sign up

Export Citation Format

Share Document