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Development of Adenosine Deaminase-Specific IgY Antibodies: Diagnostic and Inhibitory Application.

Posted by on in 2017
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Łupicka-Słowik A1Psurski M2Grzywa R1Bobrek K3Smok P1Walczak M1Gaweł A3Stefaniak T4Oleksyszyn J1Sieńczyk M5Appl Biochem Biotechnol. 2017 Oct 17. doi: 10.1007/s12010-017-2626-x. [Epub ahead of print]

1
Faculty of Chemistry, Division of Medicinal Chemistry and Microbiology, Wroclaw University of Science and Technology, Wybrzeże Wyspiańskiego 27, 50-370, Wrocław, Poland.
2
Laboratory of Experimental Anticancer Therapy, Department of Experimental Oncology, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114, Wrocław, Poland.
3
Faculty of Veterinary Medicine, Department of Epizootiology and Clinic of Bird and Exotic Animals, Wroclaw University of Environmental and Life Sciences, Pl. Grunwaldzki 45, 50-366, Wrocław, Poland.
4
Faculty of Veterinary Medicine, Department of Immunology, Pathophysiology and Veterinary Preventive Medicine, Wroclaw University of Environmental and Life Sciences, Norwida 31, 50-375, Wrocław, Poland.
5
Faculty of Chemistry, Division of Medicinal Chemistry and Microbiology, Wroclaw University of Science and Technology, Wybrzeże Wyspiańskiego 27, 50-370, Wrocław, Poland. marcin.sienczyk@pwr.edu.pl.

Abstract

Adenosine deaminase (ADA) is currently used as a diagnostic marker for tuberculous pleuritis. Although ADA has been suggested as a potential marker for several types of cancer, the importance of each of ADA isoforms as well as their levels and enzymatic activities in tumors need to be further investigated. Herein we developed avian immunoglobulin Y highly specific to human ADA via hens immunization with calf adenosine deaminase. The obtained antibodies were used for the development of a sensitive double-egg yolk immunoglobulin (IgY) sandwich ELISA assay with an ADA detection limit of 0.5 ng/ml and a linearity range of up to 10 ng/ml. Specific, affinity-purified IgYs were able to recognize human recombinant ADA and ADA present in human cancer cell lines. In addition, antigen-specific IgY antibodies were able to inhibit catalytic activity of calf ADA with an IC50 value of 47.48 nM. We showed that generated IgY antibodies may be useful for ADA detection, thus acting as a diagnostic agent in immunoenzymatic assays.

KEYWORDS:

Adenosine deaminase (ADA); Affinity purification; Anti-adenosine deaminase antibody; ELISA; Egg yolk immunoglobulin (IgY); Enzyme inhibition

PMID:
 
29043661
 
DOI:
 
10.1007/s12010-017-2626-x
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