Abstract
We applied a ribosome display technique to a mouse single chain variable fragment (scFv) library to select scFvs specific for the inducible costimulator (ICOS). mRNA was isolated from the spleens of BALB/c mice immunized with ICOS protein. Heavy and κ chain genes (VH and κ) were amplified separately by reverse transcriptase polymerase chain reaction, and the anti-ICOS VH/κ chain ribosome display library was constructed with a special flexible linker by overlap extension PCR. The VH/κ chain library was transcribed and translated in vitro using a rabbit reticulocyte lysate system. Then, antibody–ribosome–mRNA complexes were produced and panned against ICOS protein under appropriate conditions. However, in order to isolate specific scFvs for ICOS, negative selection using CD28 was carried out before three rounds of positive selection on ICOS. After three rounds of panning, the selected scFv DNAs were cloned into pET43.1a and detected by SDS-PAGE. Then, enzyme-linked immunosorbent assay showed that we successfully constructed a native ribosome display library, and among seven clones, clone 5 had the highest affinity for the ICOS and low for the CD28. Anti-ICOS scFvs are assessed for binding specificity and affinity and may provide the potential for development of the humanized and acute and chronic allograft rejection.
Similar content being viewed by others
References
Lenschow, D. J., Walunas, T. L., & Bluestone, J. A. (1996). Annual Review of Immunology, 14, 233–258.
Tamatani, T., Tezuka, K., & Hanzawa-Higuchi, N. (2000). International Immunology, 12, 51–55.
Hutloff, A., Dittrich, A. M., Beier, K. C., Eljaschewitsch, B., Kraft, R., Anagnostopoulos, I., et al. (1999). Nature, 397, 263–266.
Yoshinaga, S. K., Whoriskey, J. S., Khare, S. D., Sarmiento, U., Guo, J., Horan, T., et al. (1999). Nature, 402, 827–832.
Coyle, A. J., Lehar, S., Lloyd, C., Tian, J., Delaney, T., Manning, S., et al. (2000). Immunity, 13, 95–105.
Swallow, M. M., Wallin, J. J., & Sha, W. C. (1999). Immunity, 11, 423–432.
Rottman, J. B., Smith, T., Tonra, J. R., Ganley, K., Bloom, T., Silva, R., et al. (2001). Nature Immunology, 2, 605–611.
Ozkaynak, E., Gao, W., Shemmeri, N., Wang, C., Gutierrez-Ramos, J. C., Amaral, J., et al. (2001). Nature Immunology, 2, 591–596.
Pan, X. C., Guo, L., Deng, Y. B., Naruse, K., Kimura, H., Sugawara, Y., et al. (2008). Surgery Today, 38, 815–825.
Rando, R. F., & Notkins, A. L. (1994). Current Topics in Microbiology and Immunology, 187, 195–205.
Huston, J. S., Mudgett-Hunter, M., Tai, M. S., McCartney, J., Warren, F., Haber, E., et al. (1991). Methods in Enzymology, 203, 46–88.
Hanes, J., & Plückthun, A. (1997). Proceedings of the National Academy of Sciences, 94, 4937–4942.
He, M., & Taussig, M. J. (1997). Nucleic Acids Research, 25, 5132–5134.
He, M., & Taussig, M. J. (2005). Journal of Immunological Methods, 297, 73–82.
He, M., Menges, M., Groves, M. A., Corps, E., Liu, H., Brüggemann, M., et al. (1999). Journal of Immunological Methods, 31, 105–117.
Lee, M. S., Kwon, M. H., Kim, K. H., Shin, H. J., Park, S., & Kim, H. I. (2004). Journal of Immunological Methods, 284, 147–157.
Hanes, J., Jermutus, L., Weber-Bornhauser, S., Bosshard, H. R., & Plückthun, A. (1998). Proceedings of the National Academy of Sciences, 95, 14130–14135.
Hanes, J., Schaffitzel, C., Knappik, A., & Plückthun, A. (2000). Nature Biotechnology, 18, 1287–1292.
Yuan, Q., Xia, Y. X., Nian, S. J., Yin, Y. P., Cao, Y. Q., & Wang, Z. K. (2007). Enzyme and Microbial Technology, 41, 383–389.
Bradford, M. M. (1976). Analytical Biochemistry, 72, 248–254.
Laemmli, U. K. (1970). Nature, 277, 680–685.
Rothe, A., Nathanielsz, A., Hosse, R. J., Oberhäuser, F., Strandmann, E. P., Engert, A., et al. (2007). Journal of Biotechnology, 130, 448–454.
Kawamoto, M., Harigai, M., Hara, M., Kawaguchi, Y., Tezuka, K., Tanaka, M., et al. (2006). Arthritis Research & Therapy, 8, R62.
Nanji, S. A., Hancock, W. W., Anderson, C. C., Adams, A. B., Luo, B., Schur, C. D., et al. (2004). American Journal of Transplantation, 4, 526–536.
Nanji, S. A., Hancock, W. W., Anderson, C. C., Zhu, L. F., Kneteman, N. M., & Shapiro, A. M. (2003). Transplantation Proceedings, 35, 2477–2478.
Guo, L., Li, X. K., Funeshima, N., Fujino, M., Nagata, Y., Kimura, H., et al. (2002). Transplantation, 73, 1027–1032.
Futamatsu, H., Suzuki, J., Kosuge, H., Yokoseki, O., Kamada, M., Ito, H., et al. (2003). Cardiovascular Research, 59, 95–104.
Gonzalo, J. A., Tian, J., Delaney, T., Corcoran, J., Rottman, J. B., Lora, J., et al. (2001). Nature Immunology, 2, 597–604.
Guillonneau, C., Aubry, V., Renaudin, K., Seveno, C., Usal, C., Tezuka, K., et al. (2005). Transplantation, 80, 255–263.
Yokota, T., Milenic, D. E., Whitlow, M., & Schlom, J. (1992). Cancer Research, 52, 3402–3408.
Zhao, X. L., Chen, W. Q., Yang, Z. H., Li, J. M., Zhang, S. J., & Tian, L. F. (2009). Journal of Biotechnology, 144, 253–258.
Irving, R. A., Coia, G., Roberts, A., Nuttall, S. D., & Hudson, P. J. (2001). Journal of Immunological Methods, 248, 31–45.
Zhou, J. M., Fujita, S., Warashina, M., Baba, T., & Taira, K. (2002). Journal of the American Chemical Society, 124, 538–543.
Acknowledgments
This study was supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Pan, Y., Mao, W., Liu, X. et al. Selection of Single Chain Variable Fragments Specific for the Human-Inducible Costimulator Using Ribosome Display. Appl Biochem Biotechnol 168, 967–979 (2012). https://doi.org/10.1007/s12010-012-9800-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-012-9800-y