Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/130026
Title: Use of phage display to isolate specific human monoclonal antibody fragments against a potential target for multiple myeloma
Authors: Pei, X.L.
Ge, F.
Gullo, C. 
Teoh, G.K.H. 
Hwang, W.Y.K. 
Keywords: Antibody isolation
Ku86
Phage display
ScFv
Issue Date: Jul-2009
Citation: Pei, X.L., Ge, F., Gullo, C., Teoh, G.K.H., Hwang, W.Y.K. (2009-07). Use of phage display to isolate specific human monoclonal antibody fragments against a potential target for multiple myeloma. Annals of the Academy of Medicine Singapore 38 (7) : 621-629. ScholarBank@NUS Repository.
Abstract: Introduction: Multiple myeloma (MM), a malignancy of plasma cells, accounts for 10% of all haematological malignancies and is currently incurable. Although it can be treated, the disease tends to relapse after several years and becomes increasingly resistant to conventional therapy. Investigations into using humoral therapy for MM are now underway with a view that novel therapeutic agents may provide a more targeted therapy for MM. Materials and Methods: Here, phage display, a faster and more efficient method compared to classical hybridoma fusion technology, was used as a proof-of-concept to isolate several single-chain Fragment variables (scFv) against Ku86. Results: Anti-Ku86 polyclonal scFvs biopanning was successful where third round scFvs (A 450∼1.1) showed a 1/3 increase in binding as compared to the first round scFvs (A450∼0.4) with 100ug/mL of antigen (purified human Ku86). Subsequent selection and verification of monoclonal antibodies using third round biopanning revealed 4 good affinity binding clones ranging from A450∼0.1 to A450∼0.15 on 12.5ug/mL of antigen as compared to low binders (A450∼0.07) and these antibodies bind to Ku86 in a specific and dose-dependent manner. Comparative studies were also performed with commercially available murine antibodies and results suggest that 2 of the clones may bind close to the following epitopes aa506-541 and aa1-374. Conclusions: These studies using phage display provide an alternative and viable method to screen for antibodies quickly and results show that good affinity antibodies against Ku86 have been successfully isolated and they can be used for further studies on MM and form the basis for further development as anti-cancer therapeutic agents.
Source Title: Annals of the Academy of Medicine Singapore
URI: http://scholarbank.nus.edu.sg/handle/10635/130026
ISSN: 03044602
Appears in Collections:Staff Publications

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