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DOI: 10.5593/SGEM2016/HB63/S08.028

COMPARATIVE ANALYSIS OF POLYCLONAL AND MONOCLONAL IMMUNE PEROXIDASE CONJUGATES SPECIFIC TO BOVINE IGG FROM THE DIAGNOSTIC KITS FOR DETECTING IGG CLASS ANTIBODIES

A. K. Barsukov, A. I. Kuznetsov, O. Y. Nesterova
Friday 11 November 2016

References: 16th International Multidisciplinary Scientific GeoConference SGEM 2016, SGEM Vienna GREEN Extended Scientific Sessions, www.sgemviennagreen.org, SGEM2016 Conference Proceedings, ISBN 978-619-7105-79-7 / ISSN 1314-2704, 2 - 5 November, 2016, Book 6 Vol. 3, 215-222pp, DOI: 10.5593/SGEM2016/HB63/S08.028

ABSTRACT
In accordance with the WHO recommendations, the diagnostic tools for medical purposes based on indirect enzyme immunoassay schemes should be completed with anti-species conjugates synthesized on the polyclonal IgG fraction of rum blood immune plasma (serum). In the diagnosis of infectious animal diseases such restrictions are absent. We examined anti-species immune peroxidase conjugates specific to bovine IgG and synthesized on polyclonal and monoclonal antibodies. Taking into account all known IgG subclasses, the native cattle plasma was subjected to column rechromatography in Sephadex G-200 gel. The resulting IgG fraction was equilibrated with 0.01 mol/l sodium phosphate buffer with pH 8.0 and was subjected to anion exchange chromatography in weak anion-exchange gel using gradient elution. The IgG subfractions with different isoelectric points were formed on the basis of analytical isoelectric focusing. The subfraction samples were used to construct the calibration curves from 10 ng/ml to 10 μg/ml in the direct ELISA method.
It was found out that polyclonal immune peroxidase conjugates basically react with basic and acidic IgG, but the higher level of affinity is registered with the main IgG fraction in a comparative aspect. A similar effect characterizes the monoclonal immune peroxidase conjugates but the higher affinity reveals for the acidic IgG fraction.
We consider using the diagnostic kits including the immune enzyme conjugates of the monoclonal antibody base because their antigenic specificity takes into account the antigenic uniqueness of IgG subclasses in the formation of the optimal combination of screening tests.

Keywords: IgG subclasses, immune peroxidase conjugates.

PAPER DOI: 10.5593/SGEM2016/HB63/S08.028, COMPARATIVE ANALYSIS OF POLYCLONAL AND MONOCLONAL IMMUNE PEROXIDASE CONJUGATES SPECIFIC TO BOVINE IGG FROM THE DIAGNOSTIC KITS FOR DETECTING IGG CLASS ANTIBODIES

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