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Updated: May 8, 2026
A High-Throughput Luciferase Assay to Evaluate Proteolysis of the Single-Turnover Protease PCSK9
Published on: August 28, 2018
[Cloning and expression of human pcsk9 cDNA and preparation of its polyclonal antibody]
Fengmei Li1, Hui Yu, Jinlong Song
1Department of Clinical Laboratory, Guangzhou Medical University, Guangzhou, China.
Objective:
To construct the prokaryotic expression vector of human proprotein convertase subtilisin/kexin type 9 (pcsk9), induce it to express and prepare the polyclonal antibody of human pcsk9.
Methods:
The sequence of pcsk9 cDNA was synthesized and amplied by PCR. By means of digestion and subcloning, the sequence was cloned into the expression vector PET-28b. The recombinant plasmid of pET-28b-pcsk9 was transformed into the competent cells of E.coli BL21 (DE3), induced to express, purified and identified. The New Zealand rabbits were immunized with the purified protein. The resulted polyclonal antibody was analyzed by ELISA and Western blotting.
Results:
The prokaryotic expression vector PET-28b-pcsk9 was constructed and expressed well in E.coli BL21 (DE3). We obtained the polyclonal antibody by immunizing the rabbits with the target protein. The purified antibody was proved to have a good specificity by Western blotting and its titer reached 1:13 200.
Conclusion:
The expression vector of pcsk9 was successfully constructed and expressed. The polycolonal antibody of pcsk9 protein was obtained as well.
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