Construction of rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1369-1373, 2011.
Article
in Chinese
| WPRIM
| ID: wpr-235121
ABSTRACT
<p><b>OBJECTIVE</b>To construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.</p><p><b>METHODS</b>Peripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.</p><p><b>RESULTS</b>The libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).</p><p><b>CONCLUSION</b>The constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Arthritis, Rheumatoid
/
Recombinant Proteins
/
Immunoglobulin G
/
Molecular Sequence Data
/
Lymphocytes
/
Transfection
/
Amino Acid Sequence
/
Cloning, Molecular
/
Immunoglobulin Heavy Chains
/
Immunoglobulin kappa-Chains
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2011
Type:
Article
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