Journal of Natural Medicines

, Volume 72, Issue 1, pp 310–316 | Cite as

Different expression systems resulted in varied binding properties of anti-paclitaxel single-chain variable fragment antibody clone 1C2

  • Gorawit Yusakul
  • Seiichi Sakamoto
  • Poomraphie Nuntawong
  • Hiroyuki TanakaEmail author
  • Satoshi Morimoto


The binding properties of recombinant antibody fragments, such as affinity and specificity, determine their usefulness for therapeutic and analytical applications. Anti-paclitaxel single-chain variable fragment clone 1C2 (anti-PT scFv1C2) was expressed using Escherichia coli cell and Bombyx mori larvae expression systems. The binding characteristics of the scFvs were evaluated using indirect competitive ELISA. The linear range of binding between anti-PT scFv1C2 and paclitaxel was significantly different between the anti-PT scFv1C2s derived from E. coli (1.056–5.700 μg/ml) and B. mori (7.813–1000 ng/ml), which indicated that different expression systems resulted in different sensitivities for paclitaxel determination. In addition, the binding specificity of anti-PT scFv1C2 varied between expression systems. This finding implied that the expression system significantly affects the binding properties of recombinant antibodies, especially antibodies against low-molecular-weight targets.


Single-chain variable fragment Expression systems Specificity Analytical performance 



We acknowledge financial support from the Japan Society for the Promotion of Science (No. 22501046 and No. 19590119) and from the Takeda Science Foundation. We also appreciate the technical support from the Research Support Center, Graduate School of Medical Sciences, Kyushu University, Japan.

Compliance with ethical standards

Conflict of interest

The authors have declared no conflict of interest.

Supplementary material

11418_2017_1136_MOESM1_ESM.docx (795 kb)
Supplementary material 1 (DOCX 796 kb)


  1. 1.
    Zehnder-Fjallman AH, Marty C, Halin C et al (2007) Evaluation of anti-VEGFR-3 specific scFv antibodies as potential therapeutic and diagnostic tools for tumor lymph-angiogenesis. Oncol Rep 18:933–941Google Scholar
  2. 2.
    Safdari Y, Ahmadzadeh V, Khalili M et al (2016) Use of single chain antibody derivatives for targeted drug delivery. Mol Med 22:258CrossRefGoogle Scholar
  3. 3.
    Gould LH, Sui J, Foellmer H et al (2005) Protective and therapeutic capacity of human single-chain Fv-Fc fusion proteins against West Nile Virus. J Virol 79:14606–14613CrossRefGoogle Scholar
  4. 4.
    Turatti F, Mezzanzanica D, Nardini E et al (2001) Production and validation of the pharmacokinetics of a single-chain Fv fragment of the MGR6 antibody for targeting of tumors expressing HER-2. Cancer Immunol Immunother 49:679–686CrossRefGoogle Scholar
  5. 5.
    Maynard JA, Maassen CBM, Leppla SH et al (2002) Protection against anthrax toxin by recombinant antibody fragments correlates with antigen affinity. Nat Biotechnol 20:597–601CrossRefGoogle Scholar
  6. 6.
    Sepulveda J, Mukherjee J, Tzipori S et al (2010) Efficient serum clearance of botulinum neurotoxin achieved using a pool of small antitoxin binding agents. Infect Immun 78:756–763CrossRefGoogle Scholar
  7. 7.
    Mousli M, Devaux C, Rochat H et al (1999) A recombinant single-chain antibody fragment that neutralizes toxin II from the venom of the scorpion Androctonus australis hector. FEBS Lett 442:183–188CrossRefGoogle Scholar
  8. 8.
    Sakamoto S, Taura F, Putalun W et al (2009) Construction and expression of specificity-improved single-chain variable fragments against the bioactive naphthoquinone, plumbagin. Biol Pharm Bull 32:434–439CrossRefGoogle Scholar
  9. 9.
    Sakamoto S, Pongkitwitoon B, Nakamura S et al (2011) Construction, expression, and characterization of a single-chain variable fragment antibody against 2,4-dichlorophenoxyacetic acid in the hemolymph of silkworm larvae. Appl Biochem Biotechnol 164:715–728CrossRefGoogle Scholar
  10. 10.
    Rangnoi K, Jaruseranee N, O’Kennedy R et al (2011) One-step detection of aflatoxin-B(1) using scFv-alkaline phosphatase-fusion selected from human phage display antibody library. Mol Biotechnol 49:240–249CrossRefGoogle Scholar
  11. 11.
    Sakamoto S, Pongkitwitoon B, Nakamura S et al (2010) Efficient silkworm expression of single-chain variable fragment antibody against ginsenoside Re using Bombyx mori nucleopolyhedrovirus bacmid DNA system and its application in enzyme-linked immunosorbent assay for quality control of total ginsenosides. J Biochem 148:335–340CrossRefGoogle Scholar
  12. 12.
    Dojima T, Nishina T, Kato T et al (2010) Production of scFv-displaying BmNPV in silkworm larvae and its efficient purification. Biotechnol Appl Biochem 57:63–69CrossRefGoogle Scholar
  13. 13.
    Ishikiriyama M, Nishina T, Kato T et al (2009) Human single-chain antibody expression in the hemolymph and fat body of silkworm larvae and pupae using BmNPV bacmids. J Biosci Bioeng 107:67–72CrossRefGoogle Scholar
  14. 14.
    Kobayashi N, Oyama H (2011) Antibody engineering toward high-sensitivity high-throughput immunosensing of small molecules. Analyst 136:642–651CrossRefGoogle Scholar
  15. 15.
    Lemeulle C, Chardès T, Montavon C et al (1998) Anti-digoxin scFv fragments expressed in bacteria and in insect cells have different antigen binding properties. FEBS Lett 423:159–166CrossRefGoogle Scholar
  16. 16.
    Yusakul G, Sakamoto S, Tanaka H et al (2016) Efficient expression of single chain variable fragment antibody against paclitaxel using the Bombyx mori nucleopolyhedrovirus bacmid DNA system and its characterizations. J Nat Med 70:592–601CrossRefGoogle Scholar
  17. 17.
    Bu D, Zhou Y, Tang J et al (2013) Expression and purification of a novel therapeutic single-chain variable fragment antibody against BNP from inclusion bodies of Escherichia coli. Protein Expr Purif 92:203–207CrossRefGoogle Scholar
  18. 18.
    Weiler EW, Zenk MH (1976) Radioimmunoassay for the determination of digoxin and related compounds in Digitalis lanata. Phytochemistry 15:1537–1545CrossRefGoogle Scholar
  19. 19.
    Chao Z, Tan M, Paudel MK et al (2013) Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) using highly specific monoclonal antibody against paclitaxel. J Nat Med 67:512–518CrossRefGoogle Scholar
  20. 20.
    Vendel MC, Favis M, Snyder WB et al (2012) Secretion from bacterial versus mammalian cells yields a recombinant scFv with variable folding properties. Arch Biochem Biophys 526:188–193CrossRefGoogle Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan KK 2017

Authors and Affiliations

  • Gorawit Yusakul
    • 1
    • 2
  • Seiichi Sakamoto
    • 1
  • Poomraphie Nuntawong
    • 3
  • Hiroyuki Tanaka
    • 1
    Email author
  • Satoshi Morimoto
    • 1
  1. 1.Department of Pharmacognosy, Graduate School of Pharmaceutical SciencesKyushu UniversityFukuokaJapan
  2. 2.School of PharmacyWalailak UniversityNakhon Si ThammaratThailand
  3. 3.Faculty of Pharmaceutical SciencesChulalongkorn UniversityBangkokThailand

Personalised recommendations