In vitro selection of a peptide inhibitor of human IL-6 using mRNA display.
Interleukin-6 (IL-6) plays a crucial role in malignant diseases, such as rheumatoid arthritis, Castleman disease, and multiple myeloma, and as such, is an attractive therapeutic target. Here, the authors isolated a novel IL-6 inhibitor peptide by in vitro selection using mRNA display. The authors first used a random-primed human cDNA library to isolate IL-6-binding peptides. After four rounds of selection, a 19-amino acid peptide named CA11 was selected and confirmed to specifically interact with IL-6. The authors then performed an alanine scan analysis of CA11 and determined the amino acid residues necessary to interact with IL-6. Next, the authors constructed a CA11-based partially randomized library and after ten more rounds of selection, isolated several groups of peptides. The most frequently occurring sequence, RA07, bound to IL-6 with 3 to 4-fold higher affinity than CA11. Furthermore, RA07 inhibited IL-6-dependent KT-3 cell proliferation in a dose-dependent manner. ELISAs revealed that RA07 could not inhibit IL-6 from binding to the IL-6 receptor (IL-6R), but could inhibit the IL-6/IL-6 complex binding to gp130.
ZoeGene Corporation, 1000 Kamoshida-cho, Aoba-ku, Yokohama, Kanagawa, 227-8502, Japan. 6205732@cc.m-kagaku.co.jp
Teruaki
T
Kobayashi
Minako
M
Kakui
Tatsuro
T
Shibui
Yasunori
Y
Kitano
D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002474:Cell-Free System; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics; D010455:Peptides / Q000302:isolation & purification* / Q000494:pharmacology*; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D000093242:Interleukin-6 Inhibitors
D015850:Interleukin-6; D010455:Peptides; D012333:RNA, Messenger; D000093242:Interleukin-6 Inhibitors