Protein expression of human neuron-specific enolase and its antiserum preparation
XIAO Xin-li
HAN Jun
ZHANG Lin
CHEN Lan
ZHANG Jin
CHEN Xin-lin
ZHOU Wei
JIANG Hui-ying
ZHANG Bao-yun
LIU Yong
DONG Xiao-ping
Abstract：Objective To clone and express human neuron-specific enolase (HuNSE) protein and prepare NSE-specific antibody for prion disease diagnosis. Methods HuNSE gene was amplified by RT-PCR and subcloned into a HIS-tagged expression vector pQE30 after sequence verification. HIS-NSE fusion protein expression was obtained in E. coli M15 after IPTG induction followed by purification of the fusion protein by Ni-NTA affinity chromatography. Two male rabbits were immunized for 4 times with the purified protein, and the antiserum against NSE protein was collected and evaluated by enzyme-linked immunosorbent assay (ELISA), Western blotting and immunohischemistry. Results SDS-PAGE assay yielded an approximately 22 kD HIS-NSE fusion protein. The prepared antiserum could recognize both recombinant NSE protein and native NSE protein extracted from the brain tissues of different mammalian species as shown by Western blotting and immunohischemistry. Conclusion High expression of HuNSE is obtained in E. coli and the prepared antiserum against HuNSE can be used potentially for diagnosis ofprion-associated diseases and other nervous degeneration diseases.
Keywords：neuron-specific enolaseantiserumprion diseases
Publication Date：2006-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 1543-1547 )