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[Expression and polyclonal antibody preparation of HPV-11E7 protein].

[Article in Chinese]

Author information

1
Department of Dermatology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
2
Center of Biomedicine Research, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
3
Center of Clinical Medicine Research, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.

Abstract

OBJECTIVE:

To express human papillomavirus type 11, E7 protein (HPV11E7) via a prokaryotic expression vector and produce anti-HPV11E7 polyclonal antibody.

METHODS:

A prokaryotic expression vector pGEX-4T2-HPV11E7 was constructed and soluble GST-HPV11E7 fusion protein was expressed in E.coli by IPTG induction and purified. The purified HPV11E7 protein was used to immunize New Zealand rabbits to prepare the anti-HPV11E7 polyclonal antibody followed by protein G agarose purification to obtain the IgG type polyclonal antibody. Western blotting and immunofluorescence analysis were used to test the specificity and titer of the antibody.

RESULTS:

SDS-PAGE analysis demonstrated that large amounts of soluble GST-HPV11E7 fusion protein was expressed in E.coli after 6 hours of IPTG induction. Western blotting and immunofluorescence confirmed that the purified anti-HPV11E7 polyclonal IgG antibody obtained from immunized rabbits had a high titer and specificity.

CONCLUSION:

The prokaryotic expression system could express a great deal of soluble HPV11E7 protein, and anti-HPV11E7 polyclonal IgG antibody from HPV11E7-immunized rabbits was proved to have a high titer and specificity.

PMID:
24909284
[Indexed for MEDLINE]
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