[Expression and bioactivity analysis of the fusion protein GFP-V(L) of the neutralizing monoclonal antibody MA18/7 against HBV].
OBJECTIVE
To express the fusion protein of enhanced green fluorescent protein (EGFP) with the light chain variable domain of the neutralizing monoclonal antibody MA18/7 (mAb) against hepatitis B virus in E.coli, and determine its bioactivity.


METHODS
The EGFP gene was cloned into vector pTO-T7 to construct an expression vector. And then according to ORF gene, MA18/7-V(L) was inserted into the 5' terminal of EGFP gene free of terminal code TAA to construct expression vector of fusion protein. The fusion protein was expressed in E.coli and its bioactivity was detected with ELISA and relative fluorescence intensity.


RESULTS
The expression vector EGFP-V(L) was constructed. SDS-PAGE analysis showed that expressed fusion protein was mainly in the form of inclusion body. The fusion protein retained the property of EGFP and it could bind to V(H) to form Fv which had binding activity to pre-S1.


CONCLUSIONS
The obtained fusion protein had good bioactivity and could be applied to further studies.
Guan
Bao-quan
BQ
Ministry of Education, Key Lab of Cell Biology and Tumor Cell Engineering, Xiamen University, Xiamen 361005, China.
Zhang
Jun
J
Luo
Wen-xin
WX
Gu
Ying
Y
Zhu
Zi-heng
ZH
Xia
Ning-shao
NS
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics; D002479:Inclusion Bodies / Q000235:genetics / Q000378:metabolism; D020673:Protein Renaturation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014170:Transformation, Genetic
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins; C055966:immunoglobulin Fv; D049452:Green Fluorescent Proteins