[Expression, purification and identification of Echinococcus granulosus recombinant antigen B].
OBJECTIVE
To purify and identify recombinant antigen B of hydatid disease.


METHODS
The recombinant plasmid pMalc2x-AgB was expressed in E. coli JM109. The fusion protein rAgB-MBP was made up of antigen B and MBP (maltose binding protein) which was designed to absorb the antigen B onto the amylose column. In order to get the pure antigen as the probe for selecting phage displayed antibody library, factor Xa protease was used to digest the fusion protein rAgB-MBP so that MBP was cut off from the special cleavage site. Flowing through the amylose resin column and hydroxyapatite column, rAgB was purified by the method of affinity chromatography. Its specificity was proved by patient sera with Western blotting.


RESULTS
The recombinant antigen B was Mr 12 000, and it showed the capability to combine with the specific antibody.


CONCLUSIONS
Hydatid disease antigen B can be produced by molecular method and applied in monoclonal antibody production and phage antibody library scanning.
Surgical Lab, The First Affiliated Hospital, Medical School, Zhejiang Universtiy, Hangzhou, China.
Xin-hua
XH
Chen
Hao
H
Wen
Xiao-mei
XM
Lu
Jin-hui
JH
Zhang
Ren-yong
RY
Lin
Shu-sen
SS
Zheng
D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000378:metabolism; D002846:Chromatography, Affinity; D048209:Echinococcus granulosus / Q000276:immunology*; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D000947:Antigens, Helminth; D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins