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Expression and antigen activity determination of human thyroid peroxidase in silkworm and yeast
Authors:Baodang Guo  Wanli Gao  Huifang Lü  Xiaoliang Zhang  Xiaogui Yao  Lunguang Yao
Institution:1.Henan Provincial Engineering Laboratory of Insects Bio-reactor,Nanyang Normal University,Henan,China;2.Autobio Diagnostics Company Limited,Henan,China;3.Shaoyang Central Hospital,Hunan,China
Abstract:In this study, we report the expression of human thyroid peroxidase (TPO) in silkworm larvae and Pichia pastoris GS115. Recombinant TPO is sequentially purified from the hemolymph of infected silkworm larvae and yeast using a Ni-NTA resin kit. The concentration of yield of recombinant TPO is 4.87 mg per thousand larvae and 40.83 mg per liter yeast culture. However, the recombinant TPO produced in silkworm show similar binding ability with the specific anti-TPO serum to standard human TPO purified from insect cells. The lower antigen activity indicates the TPO expressed in yeast is not suitable to be used as the coating antigen in enzyme linked immunosorbent assay (ELISA). The cost of TPO expressed in B. mori is about 1/4 that of in insect cells, and the cost of TPO purified from silkworm for ELISA is only 1/8 that of TPO produced from Sf9 cells. It indicates the BmNPV-silkworm expression system is a cost-effective platform for producing TPO with high antigen activity.
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