Łupicka-Słowik A1, Psurski M2, Grzywa R1, Bobrek K3, Smok P1, Walczak M1, Gaweł A3, Stefaniak T4, Oleksyszyn J1, Sieńczyk M5. Appl Biochem Biotechnol. 2017 Oct 17. doi: 10.1007/s12010-017-2626-x. [Epub ahead of print]
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.
Adenosine deaminase (ADA); Affinity purification; Anti-adenosine deaminase antibody; ELISA; Egg yolk immunoglobulin (IgY); Enzyme inhibition