Stable production of recombinant chicken antibody in CHO-K1 cell line.
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.
Shimamoto
Toshi
T
Laboratory of Immunobiology, Department of Molecular and Applied Bioscience, Graduate School of Biosphere Science, Hiroshima University, 1-4-4 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8528, Japan.
Nishibori
Nahoko
N
Aosasa
Masayoshi
M
Horiuchi
Hiroyuki
H
Furusawa
Shuichi
S
Matsuda
Haruo
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001483:Base Sequence; D015153:Blotting, Western; D016466:CHO Cells; D002645:Chickens / Q000276:immunology*; D006224:Cricetinae; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D007136:Immunoglobulins / Q000096:biosynthesis*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D017931:DNA Primers; C016512:IgY; D007136:Immunoglobulins; D011994:Recombinant Proteins