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Production of Rabbit Polyclonal Antibody Using hAPOA1 Protein Expressed in Prokaryotic Cells

Production of Rabbit Polyclonal Antibody Using hAPOA1 Protein Expressed in Prokaryotic Cells
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摘要 The open reading frame (ORF) of hAPOA1 was inserted into the prokaryotic expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX- 4T-I-hAPOA1, which was then transformed into Escherichia coil strain BL21. The expression of target fusion protein was induced with isopropyl β-D-l-thiogalacto- pyranoside (IPTG). The purified fusion protein in inclusion bodies was used to immunize New Zealand white rabbits to prepare hAPOA1 antiserum and the antibody titer was detected with indirect enzyme-linked immunosorbent assay (ID-EL1SA). ID-ELISA and Western Blot proved that rabbit polyclonal antibody with a high titer of 1 : 40 000 was produced, which may bring considerable economic benefits. The open reading frame (ORF) of hAPOA1 was inserted into the prokaryotic expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX- 4T-I-hAPOA1, which was then transformed into Escherichia coil strain BL21. The expression of target fusion protein was induced with isopropyl β-D-l-thiogalacto- pyranoside (IPTG). The purified fusion protein in inclusion bodies was used to immunize New Zealand white rabbits to prepare hAPOA1 antiserum and the antibody titer was detected with indirect enzyme-linked immunosorbent assay (ID-EL1SA). ID-ELISA and Western Blot proved that rabbit polyclonal antibody with a high titer of 1 : 40 000 was produced, which may bring considerable economic benefits.
出处 《Agricultural Biotechnology》 CAS 2017年第2期41-44,共4页 农业生物技术(英文版)
基金 Supported by the Agricultural Science Independent Innovation Fund Jiangsu Province[CX(16)1326]
关键词 HAPOA1 Prokaryotie expression New Zealand white rabbits Fusion expression Polyelon~ antibody HAPOA1 Prokaryotie expression New Zealand white rabbits Fusion expression Polyelon~ antibody
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