When compared with mammalian IgG, chicken IgY is advantageous in terms of cross-reactivity. In this study, two plasmids were constructed for expression of recombinant chicken IgY derived from a chicken hybridoma. The first was for expression of the light (L) chain, and the other was for the heavy (H) chain with a histidine (His) tag at the carboxy-terminal. After transfection of recombinant chicken IgY gene into Chinese hamster ovary cells, a transfectant designated HF33 that secreted the specific antibody was selected. HF33 cells produced recombinant IgY with His tag at 10-15 microg/10(6) cells/24 h. On Western blotting analysis, the recombinant IgY was detected as one band for the H chain and two bands for the L chain. The recombinant IgY was successfully purified in a one-step procedure using a nickel-affinity resin. These results indicate that the present recombinant chicken IgY is useful for further applications.
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http://dx.doi.org/10.1016/j.biologicals.2005.05.002 | DOI Listing |
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