Construction and sequencing analysis of scFv antibody fragment derived from monoclonal antibody against norfloxacin (Nor155)

J. Mala, S. Puthong, Hiromi Maekawa, Y. Kaneko, T. Palaga, K. Komolpis, S. Sooksai

研究成果: ジャーナルへの寄稿記事

3 引用 (Scopus)

抄録

Norfloxacin belongs to the group of fluoroquinolone antibiotics which has been approved for treatment in animals. However, its residues in animal products can pose adverse side effects to consumer. Therefore, detection of the residue in different food matrices must be concerned. In this study, a single chain variable fragment (scFv) that recognizes norfloxacin antibiotic was constructed. The cDNA was synthesized from total RNA of hybridoma cells against norfloxacin. Genes encoding VH and VL regions of monoclonal antibody against norfloxacin (Nor155) were amplified and size of VH and VL fragments was 402 bp and 363 bp, respectively. The scFv of Nor155 was constructed by an addition of (Gly4Ser)3 as a linker between VH and VL regions and subcloned into pPICZαA, an expression vector of Pichia pastoris. The sequence of scFv Nor155 (GenBank No. AJG06891.1) was confirmed by sequencing analysis. The complementarity determining regions (CDR) I, II, and III of VH and VL were specified by Kabat method. The obtained recombinant plasmid will be useful for production of scFv antibody against norfloxacin in P. pastoris and further engineer scFv antibody against fluoroquinolone antibiotics.

元の言語英語
ページ(範囲)69-76
ページ数8
ジャーナルJournal of Genetic Engineering and Biotechnology
15
発行部数1
DOI
出版物ステータス出版済み - 6 1 2017

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Single-Chain Antibodies
Norfloxacin
Immunoglobulin Fragments
Complementarity Determining Regions
Monoclonal Antibodies
Fluoroquinolones
Anti-Bacterial Agents
Pichia
Nucleic Acid Databases
Hybridomas
Plasmids
Complementary DNA
RNA
Food
Genes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Genetics

これを引用

Construction and sequencing analysis of scFv antibody fragment derived from monoclonal antibody against norfloxacin (Nor155). / Mala, J.; Puthong, S.; Maekawa, Hiromi; Kaneko, Y.; Palaga, T.; Komolpis, K.; Sooksai, S.

:: Journal of Genetic Engineering and Biotechnology, 巻 15, 番号 1, 01.06.2017, p. 69-76.

研究成果: ジャーナルへの寄稿記事

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AU - Maekawa, Hiromi

AU - Kaneko, Y.

AU - Palaga, T.

AU - Komolpis, K.

AU - Sooksai, S.

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