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http://dx.doi.org/10.5483/BMBRep.2011.44.4.244

Ex12 helper phage improves the quality of a phage-displayed antibody library by ameliorating the adverse effect of clonal variations  

Choi, Hyo-Jung (Department of Systems Immunology, College of Biomedical Science, Kangwon National University)
Song, Suk-Yoon (IG Therapy Co., Rm A202, Biomedical Science Building, Kangwon National University)
Yoon, Jae-Bong (Department of Systems Immunology, College of Biomedical Science, Kangwon National University)
Liu, Li-Kun (Department of Systems Immunology, College of Biomedical Science, Kangwon National University)
Cho, Jae-Youl (Institute of Antibody Research, Kangwon National University)
Cha, Sang-Hoon (Department of Systems Immunology, College of Biomedical Science, Kangwon National University)
Publication Information
BMB Reports / v.44, no.4, 2011 , pp. 244-249 More about this Journal
Abstract
The quality of a phage-displayed antibody library deteriorates with clonal variations, which are caused by differentially expressed Escherichia coli antibody genes. Using the human Fab SP114 against the pyruvate dehydrogenase complex-E2 (PDCE2), we created four E. coli TOP10F' clones with a pCMTG phagemid encoding Fab-pIII (pCMTG-Fab), Fd ($V_H+C_{H1}$)-pIII (pCMTG-Fd), or light chain (L) (pCMTG-L), or the vector only (pCMTG-${\Delta}Fab$) to investigate the effect of clonal variations in a defined manner. Compared to the others, the E. coli clone with pCMTG-Fab was growth retarded in liquid culture, but efficiently produced phage progenies by Ex12 helper phage superinfection. Our results suggest that an antibody library must be cultured for a short duration before helper phage superinfection, and that the Ex12 helper phage helped to alleviate the detrimental effect of clonal variation, at least in part, by preferentially increasing functional phage antibodies during phage amplification.
Keywords
Clonal variation; Ex12 helper phage; M13KO7 helper phage; pCMTG; Phage display library;
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1 O'Connell, D., Becerril, B., Roy-Burman, A., Daws, M. and Marks, J. D. (2002) Phage versus phagemid libraries for generation of human monoclonal antibodies. J. Mol. Biol. 321, 49-56.   DOI   ScienceOn
2 Knappik, A. and Plückthun, A. (1995) Engineered turns of a recombinant antibody improve its in vivo folding. Protein Eng. 8, 81-89.   DOI
3 Cabilly, S. (1989) Growth at sub-optimal temperatures allows the production of functional, antigen-binding Fab fragments in Escherichia coli. Gene 85, 553-557.   DOI   ScienceOn
4 De Bruin, R., Spelt, K., Mol, J., Koes, R. and Quattrocchio, F. (1999) Selection of high-affinity phage antibodies from phage display libraries. Nat. Biotechnol. 17, 397-399.   DOI   ScienceOn
5 Kretzschmar, T., Zimmermann, C. and Geiser, M. (1995) Selection procedures for nonmatured phage antibodies: a quantitative comparison and optimization strategies. Analytical. Biochem. 224, 413-419.   DOI   ScienceOn
6 Mutuberria, R., Hoogenboom, H. R., VanderLinden, E., DeBruine, A. P. and Roovers, R. C. (1999) Model systems to study the parameters determining the success of phage antibody selection on complex antigens. J. Immunol. Methods 231, 65-81.   DOI   ScienceOn
7 Baek, H., Suk, K., Kim, Y. and Cha, S. (2002) An improved helper phage system for efficient isolation of specific antibody molecules in phage display. Nucleic Acids Res. 30(5), e18.   DOI   ScienceOn
8 Cha, S., Leung, P. S. C., Coppel, R. L., VandeWater, J., Ansari, A. A. and Gershwin, M. E. (1994) Heterogeneity of combinatorial human autoantibodies against PDC-E2 and biliary epithelial cells in patients with primary biliary cirrhosis. Hepatology 20, 574-583.   DOI   ScienceOn
9 McCafferty, J. (1996) Phage display: factors affecting panning efficiency; in Phage Display of Peptides and Proteins, A Laboratory Manual, pp. 261, Academic Press Inc., San Diego, USA.
10 Corisdeo, S. and Wang, B. (2004) Functional expression and display of an antibody Fab fragment in Escherichia coli: study of vector designs and culture conditions. Prot. Exp. Purif. 34, 270-279.   DOI   ScienceOn
11 Assazy, H. M. E. and Highsmith, W. E. (2002) Phage display technology: clinical applications and recent Innovations. Clin. Biochem. 35, 425-445.   DOI   ScienceOn
12 Yamanaka, H. I., Kirii, Y. and Ohmoto, H. (1995) An improved phage display antibody cloning system using newly designed PCR primers optimized for Pfu DNA polymerase. J. Biochem. (Tokyo) 117, 1218-1227.   DOI
13 Krebber, A., Bornhauser, S., Burmester, J., Honegger, A., Willuda, J., Bosshard, H. R. and Pluckthun, A. (1997) Reliable cloning of functional antibody variable domains from hybridomas and spleen cell repertoires employing a reengineered phage display system. J. Immunol. Methods 201, 35-55.   DOI   ScienceOn
14 Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning. A laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
15 Cha, S., Leung, P. S. C., Gershwin, M. E., Fletcher, M. P. and Ansari, A. A. (1993) Combinatorial autoantibodies to dihydrolipoamide acetyltransferase, the major autoantigen of primary biliary cirrhosis. Proc. Natl. Acad. Sci. 90, 2527-2531.   DOI
16 McCafferty, J. and Johnson, K. S. (1996) Construction and screening of antibody display libraries; in Phage Display of Peptides and Proteins. A Laboratory Manual. pp. 79, Academic Press Inc., San Diego, USA.
17 Ostermeier, M. and Benkovic, S. J. (2000) A two-phagemid system for the creation of non-phage displayed antibody libraries approaching one trillion members. J. Immunol. Meth. 237,175-186.   DOI   ScienceOn
18 Barbs, C. F. 3rd., Kang, A. S., Lerner, R. A. and Benkovic, S. J. (1991) Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc. Natl. Acad. Sci. 88, 7978-7982.   DOI   ScienceOn
19 Hoogenboom, H. R., Griffiths, A. D., Johnson, K. S., Chiswell, D. J., Hudson, P. and Winter, G. (1991) Multi- subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains. Nucleic Acids Res. 19, 4133-4137.   DOI   ScienceOn
20 Clark, M. A., Hammond, F. R., Papaioannou, A., Hawkins, N. J. and Ward, R. L. (1997) Regulation and expression of human Fabs under the control of the Escherichia coli arabinose promoter, PBAD. Immunotechnol. 3, 217-226.   DOI   ScienceOn
21 Dziegiel, M., Nielsen, L. K., Andersen, P. S., Blancher, A., Dickmeiss, E. and Engberg, J. (1995) Phage display used for gene cloning of human recombinant antibody against the erythrocyte surface antigen, rhesus D. J. Immunol. Methods 182, 7-19.   DOI   ScienceOn
22 Marks, J. D. and Bradbury, A. (2004) Selection of human antibodies from phage display libraries. Methods Mol. Biol. 248, 161-176.
23 Hoogenboom, H. R. and Chames, P. (2000) Natural and designer binding sites made by phage display technology. Immunol. Today 21, 371-378.   DOI   ScienceOn
24 Oh, M., Joo, H., Hur, B., Jeong, Y. and Cha, S. (2007) Enhancing phage display of antibody fragments using gIII-amber suppression. Gene 386, 81-89.   DOI   ScienceOn