Characterization of 11-dehydro-thromboxane B2 recombinant antibody obtained by phage display technology

Lilian Rumi Tsuruta, Yoshihisa Tomioka, Takanori Hishinuma, Yoshinori Kato, Kunihiko Itoh, Toshio Suzuki, Hiroki Oguri, Masahiro Hirama, Junichi Goto, Michinao Mizugaki

研究成果: Article査読

8 被引用数 (Scopus)

抄録

Recombinant monoclonal antibodies specific for 11-dehydro-thromboxane B2 (11D-TX) were isolated from the combinatorial libraries on a pComb3 phage-display vector using a magnetic cell sorting (MACS) system. The libraries were constructed from repertories of light and heavy-chains derived from the total RNA of 11D-TX conjugated keyhole limpet haemocyanin-immunized mice. Biotinylation of 11D-TX conjugated bovine serum albumin (BSA) was performed through free thiol groups on BSA using 1-biotinamido-4-[4′-(maleimidomethyl) cyclohexanecarboxamido] butane (Biotin-BMCC). Affinity bio-panning was performed to enrich the phage display libraries against biotinylated 11D-TX conjugated BSA with the MACS system. Results indicated that the selected anti-11D-TX Fab fragments expressed by E. coli exhibited a five-fold higher affinity for BSA conjugated 11D-TX compared to BSA alone and little specificity to other related compounds as determined by the binding assay and inhibition enzyme-linked immunosorbent assay (ELISA). This is the first report of an antibody against prostaglandin produced by phage display technology and also determination of the DNA sequence of this antibody. The MACS system was shown to be a simpler and more efficient method of panning than the conventional ELISA procedure. According to our results, we concluded that the phage display technique combined with the MACS system allowed the selection of the antibody with high affinity and some specificity.

本文言語English
ページ(範囲)273-284
ページ数12
ジャーナルProstaglandins Leukotrienes and Essential Fatty Acids
68
4
DOI
出版ステータスPublished - 2003 4 1

ASJC Scopus subject areas

  • 臨床生化学
  • 細胞生物学

フィンガープリント

「Characterization of 11-dehydro-thromboxane B<sub>2</sub> recombinant antibody obtained by phage display technology」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル