| TECHNIQUES AND METHODS |
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| Construction and Verification of a Novel System for B-cell Epitope Display |
| LIU Ying-fu, HUO Jing-rui, FAN Guo-cai, SUN Zhi-xian, WANG Qing-ming |
| State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China |
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Abstract B cell epitopes are the sites of the molecules that are recognized by antibodies or B cell receptors. Prediction and verification of B cell epitopes of antigen is useful for specific antibody generation and vaccine design. The immunoglobulin heavy chain variable region was used as the scaffold protein for B cell epitope display, and its CDR3 region was the display site. Three B cell epitopes were displayed in the protein scaffold, including (His)6 tag and two epitopes derived from VEGF. The three recombinant proteins were used to immunize mice, and the immune serum was isolated and analyzed by ELISA, Western blot and immunofluorescence. The results demonstrated that the immune serum exhibited good specificity for the corresponding epitopes and the source protein. This means that the immunoglobulin heavy chain variable region is a good scaffold protein for B cell epitope display.
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Received: 15 February 2012
Published: 25 June 2012
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[1] Wiesmuller K H, Fleckenstein B, Jung G. Peptide vaccines and peptide libraries. Biol Chem, 2001, 382(4):571-579.
[2] Zauner W, Lingnau K, Mattner F, et al. Defined synthetic vaccines. Biol Chem, 2001, 382(4):581-595.
[3] Jameson B A,Wolf H. The antigenic index: a novel algorithm for predicting antigenic determinants. Comput Appl Biosci, 1988, 4(1):181-186.
[4] Saha S Raghava. Prediction of continuous B-cell epitopes in an antigen using recurrent neural network. Proteins, 2006, 65(1):40-48.
[5] 万涛,孙涛,吴加金.蛋白顺序性抗原决定簇的多参数综合预测.中国免疫学杂志,1997,13(3):329-333. Wan T, Sun T, Wu J J. The multi-parameter prediction of potential protein antigenic determinants. Chinese Journal of Immunology,1997, 13(3):329-333.
[6] Markus N, Birgit W, Marc B, et al. Development of a vaccine marker technology: Display of B cell epitopes on the surface of recombinant polyomavirus-likepentamers and capsoids induces peptide-specific antibodies in piglets after vaccination. Biotechnol J, 2006, 1:1435-1446.
[7] Greenbaum J A, Andersen P H, Blythe M, et al. Towards a consensus on datasets and evaluation metrics for developing B-cell epitope prediction tools. J Mol Recognit, 2007, 20(2):75-82.
[8] Vu K B, Ghahroudi M A. Comparison of llama VH sequences from conventional and heavy chain antibodies. Mol Immunol, 1997, 34(16-17):1121-1131.
[9] Lindemann J. Speculations on idiotypes and homobodies. Ann Immunol,1973, 124(2):171-184.
[10] Jerne N K. Towards a network theory of the immune system. Ann Immunol, 1974, 125C(1-2):373-389.
[11] Maurizio Z, Frangoise R, Paola L, et al. Theoretical and practical aspects of antigenized antibodies. Immunological Reviews, 1992, 130(1):125-150.
[12] Spangler B D. Binding to native proteins by antipeptide monoclonal antibodies. J Immunol, 1991, 146(5):1591-1595.
[13] Newmark P. Will peptides make vaccines. Nature, 1983, 305(5929):9-9.
[14] Rosario B, Michael R. Immunogenicity of an engineered internal image antibody. PNAS, 1991, 88:4713-4717. |
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