Cloning and expressing of tissue inhibitor of metalloproteinases I gene fragment and preparation of monoclonal antibodies against the recombinant protein / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 231-233, 2013.
Article
in Chinese
| WPRIM
| ID: wpr-318055
ABSTRACT
<p><b>OBJECTIVE</b>To prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein.</p><p><b>METHODS</b>TIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot were used to detect specificity of mAbs.</p><p><b>RESULTS</b>The prokaryotic plasmid expressing the recombinant protein was constructed, and the TIMP-I recombinant protein was expressed and purified. Four hybridoma cell lines that secreted anti-TIMP-I mAbs were obtained. 3 of 4 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with TIMP-I protein.</p><p><b>CONCLUSION</b>The TIMP-I recombinant protein is highly purified and has strong antigenicity. The anti- TIMP-I mAbs were prepared successfully.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Proteins
/
Cloning, Molecular
/
Tissue Inhibitor of Metalloproteinase-1
/
Allergy and Immunology
/
Genetics
/
Mice, Inbred BALB C
/
Antibodies, Monoclonal
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2013
Type:
Article
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