Related Experiment Video
Updated: Jun 20, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
[Expression, purification and polyclonal antibody preparation for a novel gene BC097361]
Qi Wang1, Yun-lei Liang, Hong-shan Wei
1Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100011, China.
Objective:
To express and purify of the BC097361 recombinant protein, and to prepare the BC097361 specific rabbit polyclonal antibody.
Methods:
BC097361 cDNA was ligated into the prokaryotic expressive vector pET-32a (+), and the resulting plasmid was transformed into E.coli BL21 (DE3). The protein expression was induced with IPTG and the protein was analyzed with SDS-PAGE and western blotting. The expressed product was purified using Ni+ affinity column chromatography.Then the purified pET-32a (+) -BC097361 fusion protein was used to immunize New Zealand rabbits to gain polyclonal antibody. The specificity and potency of polyclonal antibody were evaluated by Western blot and ELISA.
Results:
The BC097361 fusion protein was highly expressed.The protein was mainly in the inclusion body. ELISA indicated the titer of polyclonal antibody more than 1:320000. The high specificity was confirmed with Western blot.
Conclusions:
The recombinant BC097361 fusion protein and the BC097361 specific polyclonal antibody will be valuable tools for the investigation on the biological function of BC097361.
