Marshall Digital Scholar

Title

Use of Phage Display to Isolate Specifi c Human Monoclonal Antibody Fragments Against a Potential Target for Multiple Myeloma

Authors

Pei Xiong Liew
Feng Ge
Charles A. Gullo, gullo@marshall.eduFollow
Gerrard KH Teoh
William YK Hwang

Document Type

Article

Publication Date

7-2009

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 (A450~1.1) 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 verifi cation of monoclonal antibodies using third round biopanning revealed 4 good affi nity 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 specifi c 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.

Comments

The copy of record is available from Annals of the Academy of Medicine, Singapore at http://www.annals.edu.sg/pdf/38VolNo7Jul2009/V38N7p621.pdf.

Copyright © 2009 Academy of Medicine, Singapore. Reprinted with permission. All rights reserved.

Recommended Citation

Liew PX, Ge F, Gullo C, Teoh GK, Hwang WY. Use of phage display to isolate specific human monoclonal antibody fragments against a potential target for multiple myeloma. Ann Acad Med, Singapore. 2009 ;38(7):621-9.