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Development of monoclonal antibodies for detection of Antisecretory Factor activity in human plasma.

Journal article
Authors Ewa Johansson
Ivar Lönnroth
Ingela Jonson
Stefan Lange
Eva Jennische
Published in Journal of immunological methods
Volume 342
Issue 1-2
Pages 64-70
ISSN 1872-7905
Publication year 2009
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Institute of Biomedicine, Department of Infectious Medicine
Pages 64-70
Language en
Links dx.doi.org/10.1016/j.jim.2008.11.01...
Keywords Adult, Animals, Antibodies, Monoclonal, analysis, biosynthesis, Antibody Specificity, Diet, Enzyme-Linked Immunosorbent Assay, methods, Female, Food, Fortified, Humans, Hybridomas, Immunoglobulin Isotypes, analysis, Male, Mice, Mice, Inbred BALB C, Middle Aged, Neuropeptides, blood, isolation & purification, Placenta, Pregnancy, Rats
Subject categories Cell and Molecular Biology, Microbiology in the medical area

Abstract

Antisecretory Factor (AF) is expressed in most tissues and can be demonstrated in plasma and other body fluids. Most of the AF in plasma is in an inactive form and activation of AF occurs after exposure to bacterial toxins or after intake of various dietary components. Patients with chronic diseases involving disturbances in inflammatory and secretory processes may benefit from an AF-inducing diet. The aim of the present study was to develop an in vitro assay for the analysis of AF-activity in human plasma. Monoclonal antibodies were raised against a native form of AF prepared from human placenta. Nine clones of the monoclonal antibodies recognizing AF and AF peptides were identified. With the aid of these antibodies, we developed a sensitive ELISA method for direct detection of AF-activity in human plasma. The AF activity in plasma from five healthy volunteers was low, 0.112+/-0.022 (absorbance at 405 nm), before intake of the AF-inducing diet with the SPC-Flakes, and increased significantly (p<0.05) to 0.444+/-0.068 after >or=6 weeks on the diet. A comparison of the plasma-AF values, obtained by the bioassay and the immunogenic assay (indirect ELISA), shows that there is a significant correlation (r=0.85) between the values from the two methods. The results indicate that the ELISA measures AF-activity and has the potential to be an important tool for the analysis of AF-activity in further clinical studies on AF-therapy.

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