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

Journal of Immunological Methods
Yoko NagumoI Kumagai

Abstract

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 K(d) approximately 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.

References

Mar 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·M J Casadaban
Aug 5, 1992·Journal of Molecular Biology·R E HawkinsG Winter
Oct 1, 1991·Protein Engineering·C CagnonJ M Masson
Sep 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·C F BarbasS J Benkovic
Aug 15, 1991·Nature·T ClacksonG Winter
Jan 1, 1990·Journal of Clinical Laboratory Analysis·Y Hokama
Jan 1, 1980·Journal of Immunological Methods·J W Goding
Apr 28, 1995·Journal of Molecular Biology·P D JeffreyS Sheriff
Jun 21, 1994·Proceedings of the National Academy of Sciences of the United States of America·H MiyashitaI Fujii
Nov 20, 1998·Biological Chemistry·S J McConnellD G Spinella
Feb 15, 2000·Toxicon : Official Journal of the International Society on Toxinology·S PauillacA M Legrand
Aug 11, 2000·Toxicon : Official Journal of the International Society on Toxinology·R J Lewis
Jun 1, 2001·Nature Biotechnology·N TakahashiI Fujii
Dec 1, 2001·Science·M HiramaM Satake
Mar 16, 2002·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·T Yasumoto
Dec 6, 2002·Organic Letters·Masayuki InoueMasahiro Hirama
Mar 12, 2003·Prostaglandins, Leukotrienes, and Essential Fatty Acids·Lilian Rumi TsurutaMichinao Mizugaki
Jun 19, 2003·Journal of the American Chemical Society·Hiroki OguriYoko Nagumo

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Citations

May 12, 2009·Seminars in Cell & Developmental Biology·Paul J ConroyRichard J O'Kennedy
Nov 28, 2007·Biochemical and Biophysical Research Communications·Takamitsu HattoriIzumi Kumagai
May 12, 2012·Toxicon : Official Journal of the International Society on Toxinology·Takeshi TsumurayaMasahiro Hirama
Jan 5, 2007·Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan·Ikuo Fujii
Oct 12, 2016·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Masahiro Hirama
May 1, 2007·Organic Letters·J Stephen ClarkAlexander J Blake
Aug 10, 2020·Ecotoxicology and Environmental Safety·G GaianiM Campàs
May 18, 2018·Analytical Chemistry·Takeshi TsumurayaIkuo Fujii

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