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

Łupicka-Słowik, Agnieszka, Psurski, Mateusz, Grzywa, Renata, Bobrek, Kamila, Smok, Patrycja, Walczak, Maciej, Gaweł, Andrzej, Stefaniak, Tadeusz, Oleksyszyn, Józef, Sieńczyk, Marcin
Applied biochemistry and biotechnology 2018 v.184 no.4 pp. 1358-1374
adenosine, adenosine deaminase, antibodies, calves, catalytic activity, enzyme activity, enzyme-linked immunosorbent assay, hens, human cell lines, humans, immunization, immunoglobulin Y, inhibitory concentration 50, neoplasms, pleurisy
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.