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...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 Bm NPV-silkworm expression system is a cost-effectiv e platform for producing TPO with high antigen activity.展开更多
基金Supported by the National Natural Science Foundation of China(31372381)Henan Science and Technology Innovation Team of Insect Bio-reactor Fund(C20140047)Henan Natural Science Foundation Project(132300413214.0)
文摘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 Bm NPV-silkworm expression system is a cost-effectiv e platform for producing TPO with high antigen activity.
