Production and characterisation of a novel chicken IgY antibody raised against C-terminal peptide from human thymidine kinase 1

Egg yolk is a good source of highly specific antibodies against mammalian antigens because of the phylogenetic distance between birds and mammals. Chicken egg yolk immunoglobulins (IgY) were generated to a synthetic 31-amino acid peptide from the C-terminal of human HeLa thymidine kinase 1 (TK1) enzyme. The anti-TK1 IgY antibody was purified using affinity chromatography against the 31-amino acid peptide. The purified antibody inhibited the catalytic activity of the TK1 enzyme in the CEM TK1 + cells and recognized the 25-kDa subunit and tetrameric form of TK1, which has a p I value of 8.3. No immunoreaction was observed in CEM TK1 − cells. Western blot of the serum TK1 (S-TK1) also showed that only a single band was found in the serum of patients with malignancies. No band was seen in healthy serum. Furthermore, dot blots and enhanced chemiluminescence (ECL) detection of S-TK1 performed on sera of preoperative patients with gastric cancer (GC) ( n=31) and healthy controls ( n=62) showed that the levels of S-TK1 in the sera of cancer patients were significantly different ( P