tetano
Editor, Senior Moderator
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2011 May;27(5):531-534.
[Purification of Cyanovirin-N with antiviral activity and preparation as well as modification of its polyclonal antibody.]
[Article in Chinese]
Han B, Yang H, Wang QL, Chen W, Qian CW, Xiong S.
Source
Biomedical Research and Development Center, Guangdong Provincial Key Laboratory of Bioengineering Medicine, National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou 510630, China.
Abstract
AIM:
To purify the recombinant Cyanovirin-N (CVN) and determine its anti-influenza virus A (H1N1) activity, and to prepare the polyclonal antibody of CVN, purify it and label it with an enzyme for future application.
METHODS:
The recombinant CVN were rapidly purified by two rounds of Ni-NTA chelating chromatography intervened with a SUMO protease cleavage step. Anti-H1N1 activity of CVN was determined using cytopathogenic effect assay (CPE). The rabbits were immunized with the purified CVN and the antibody was identified by ELISA and Western blot. IgG was purified by ammonium sulfate precipitation followed by DEAE chromatography and the purified IgG was labeled by HRP.
RESULTS:
The purity of the obtained CVN protein which showed obvious Anti-H1N1 activity in vitro was higher than 95%.The polyclonal antibody of CVN was successfully produced in the rabbits and the results of ELISA and Western blot showed that the antiserum had high titer and high specificity. The purified antibody with a titer up to 1:6 400 was successfully obtained and the anti-CVN antibody-HRP conjugate was achieved after labeling the purified antibody with HRP.
CONCLUSION:
The purified antibody against CVN has been purified and further coniugated with HRP, with can be used for future research.
PMID:
21557911
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21557911
[Purification of Cyanovirin-N with antiviral activity and preparation as well as modification of its polyclonal antibody.]
[Article in Chinese]
Han B, Yang H, Wang QL, Chen W, Qian CW, Xiong S.
Source
Biomedical Research and Development Center, Guangdong Provincial Key Laboratory of Bioengineering Medicine, National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou 510630, China.
Abstract
AIM:
To purify the recombinant Cyanovirin-N (CVN) and determine its anti-influenza virus A (H1N1) activity, and to prepare the polyclonal antibody of CVN, purify it and label it with an enzyme for future application.
METHODS:
The recombinant CVN were rapidly purified by two rounds of Ni-NTA chelating chromatography intervened with a SUMO protease cleavage step. Anti-H1N1 activity of CVN was determined using cytopathogenic effect assay (CPE). The rabbits were immunized with the purified CVN and the antibody was identified by ELISA and Western blot. IgG was purified by ammonium sulfate precipitation followed by DEAE chromatography and the purified IgG was labeled by HRP.
RESULTS:
The purity of the obtained CVN protein which showed obvious Anti-H1N1 activity in vitro was higher than 95%.The polyclonal antibody of CVN was successfully produced in the rabbits and the results of ELISA and Western blot showed that the antiserum had high titer and high specificity. The purified antibody with a titer up to 1:6 400 was successfully obtained and the anti-CVN antibody-HRP conjugate was achieved after labeling the purified antibody with HRP.
CONCLUSION:
The purified antibody against CVN has been purified and further coniugated with HRP, with can be used for future research.
PMID:
21557911
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21557911