[Construction of full-length human bladder cancer-specific antibody libraries based on mammalian display technology].

Abstract

OBJECTIVE:

To construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.

METHODS:

The total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.

RESULTS:

The libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).

CONCLUSION:

We have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.

• Authors: Kaijian Lan, Zhehuan Zhang, Zhongkun Liang, Junjie Wang, Haibo Lou, Yuanping Zhou, Shuwen Liu, Changzheng Li, Wanlong Tan, Chen Zhou
• Journal: Nan fang yi ke da xue xue bao = Journal of Southern Medical University (Nan Fang Yi Ke Da Xue Xue Bao) Vol. 33 Issue 5 Pg. 684-91 (May 2013) ISSN: 1673-4254 [Print] China
• PMID: 23688987 (Publication Type: English Abstract, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• Immunoglobulin Heavy Chains
• Immunoglobulin kappa-Chains
• Peptide Library

Topics

• Amino Acid Sequence
• Animals
• Antibodies (genetics)
• Cell Surface Display Techniques
• Gene Library
• Humans
• Immunoglobulin Heavy Chains (genetics)
• Immunoglobulin kappa-Chains (genetics)
• Peptide Library
• Urinary Bladder Neoplasms (genetics, immunology)