title: The comparison of BLyS-binding peptides from phage display library and computer-aided design on BLyS-TACI interaction.
abstract: BLyS antagonists have become the therapeutic reagents in the treatment of autoimmune disorders. BLyS binding peptides and their Fc fusion proteins may be alternative BLyS antagonists in such application. In this study, the activity of BLyS binding peptide 814 obtained from phage display library and peptide TA designed by computer-aided modeling on the interaction of BLyS-TACI was compared. In addition, to maintain the spatial conformation and stability of the peptides, human IgG1 Fc fragment was fused to peptides 814 and TA to form peptide-Fc fusion proteins, steady and innovative peptibodies. The prokaryotic expression plasmids pET30a-814-Fc and pET30a-TA-Fc for these peptibodies were acquired by genetic engineering, and confirmed by DNA sequencing. After the right plasmids were transformed into Escherichia coli BL21 (DE3), the fusion proteins were expressed and purified by protein A affinity column. As a result of competitive ELISA, peptides 814 and TA at 100μg/ml displayed 52.2% and 28.6% inhibition on the interaction of TACI-Fc with BLyS respectively. Moreover, 814-Fc and TA-Fc fusion proteins could bind to BLyS in a dosage-dependent manner as TACI-Fc did, and displayed 54.7% and 26.1% inhibition on the interaction of TACI-Fc-Myc with BLyS at 100μg/ml respectively. So 814-Fc and TA-Fc proteins had the similar bioactivity as the peptides did. Furthermore, compared with TA-Fc, 814-Fc showed two-fold inhibition effect on BLyS binding to TACI, suggesting that 814-Fc could inhibit BLyS bioactivity significantly and might serve as a potential antagonist to treat autoimmune diseases associated with BLyS overexpression.
authors.affiliation: Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, PR China.
authors.forename: Yacong
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, PR China.
authors.forename: Xiafei
authors.initials: X
authors.lastname: Hao
authors.affiliation: Institute of Basic Medical Sciences, Beijing 100850, PR China.
authors.forename: Jiannan
authors.initials: J
authors.lastname: Feng
authors.affiliation: Institute of Basic Medical Sciences, Beijing 100850, PR China.
authors.forename: Beifen
authors.initials: B
authors.lastname: Shen
authors.affiliation: Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, PR China. Electronic address: betty_wj@tju.edu.cn.
authors.forename: Jing
authors.initials: J
authors.lastname: Wei
authors.affiliation: Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, PR China. Electronic address: jsunbio2009@yahoo.com.
authors.forename: Jian
authors.initials: J
authors.lastname: Sun
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology / Q000627:therapeutic use*; D001327:Autoimmune Diseases / Q000188:drug therapy*; D053264:B-Cell Activating Factor / Q000037:antagonists & inhibitors / Q000378:metabolism*; D017076:Computer-Aided Design; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D019151:Peptide Library; D011485:Protein Binding; D011487:Protein Conformation; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000235:genetics; D053303:Transmembrane Activator and CAML Interactor Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D000893:Anti-Inflammatory Agents; D053264:B-Cell Activating Factor; D007141:Immunoglobulin Fc Fragments; D010446:Peptide Fragments; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; C108467:TNFRSF13B protein, human; D053303:Transmembrane Activator and CAML Interactor Protein
keywords: Autoimmune diseases; BLyS; BLyS antagonists; Computer-aided design; Peptibody