Phage-display selection of antibodies to the left end of CTX3C using synthetic fragments

Yoko Nagumo, Hiroki Oguri, Kouhei Tsumoto, Yumi Shindo, Masahiro Hirama, Takeshi Tsumuraya, Ikuo Fujii, Yoshihisa Tomioka, Michinao Mizugaki, Izumi Kumagai

研究成果: Article査読

21 被引用数 (Scopus)

抄録

Ciguatoxins (CTXs) are a family of toxins that contaminate a variety of fish and cause ciguatera seafood poisoning. The limited availability of CTXs from natural sources has hampered preparation of antibodies and, thus, the development of immunoassays for these toxins. In the current studies, we utilized synthetic fragments as haptens to prepare antibodies against CTX3C, a congener of CTXs, thereby avoiding the problem of its scarcity. Synthetic ABC-ring fragment (ABC) of CTX3C was conjugated with keyhole limpet hemocyanin (KLH) and immunized on mice. Phage-displayed antibodies were first screened based on affinity to a soluble biotin-linked ABC-ring fragment that was captured on streptavidin-linked magnetic beads. The beads were then eluted with the ABCD-ring fragment (ABCD), and recovered phages were amplified. This elution with a synthetic fragment allowed the preparation of antibodies to ABCD as well as to ABC. Three antibodies with affinities of Kd∼10-5 M for ABCD were selected and prepared as soluble recombinant Fabs (rFabs). One rFab, 1C49, bound to CTX3C itself, although the binding affinity for CTX3C was weaker than for ABCD.

本文言語English
ページ(範囲)137-146
ページ数10
ジャーナルJournal of Immunological Methods
289
1-2
DOI
出版ステータスPublished - 2004 6

ASJC Scopus subject areas

  • 免疫アレルギー学
  • 免疫学

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