Preparation and Characterization of Monoclonal Antibodies Suitable for Detection of Foodborne Pathogens by Biosensor

Xiao Guang Zhang, Sachiko Tsuji, Hayato Kitaoka, Mitsuru Tamai, Hiroshi Kobayashi, Ken Ichi Honjoh, Takahisa Miyamoto

Research output: Contribution to journalArticlepeer-review

Abstract

Monoclonal antibodies (MAbs) for detection of Escherichia coli O157:H7 (O157:H7), Salmonella Enteritidis (SE) and Listeria monocytogenes (LM) using surface plasmon resonance (SPR) biosensor were prepared and characterized. Indirect enzyme–linked immunosorbent assay (ELISA) and SPR biosensor were used for screening of the hybridoma cells secreting MAbs specific to the pathogens. Based on the reactivity of MAbs against the target pathogens by SPR biosensor, MAbs were selected. For O157:H7, the clones 3–11B–3F–8 and 3–11B–3F–11, which culture supernatants reacted strongly with boiled O157:H7 and sonicated O157:H7 cells were obtained and their culture supernatants were used for purification of anti–O157:H7 MAb. For SE, the clone 1–11G–8, which generated high response to sonicated SE cells and the lowest response to the sonicated mixture cells was obtained and the culture supernatant was used for purification of anti–SE MAb for detection of sonicated SE cells. The clone of 3–5H–3F was found with high reactivity against boiled SE and very low against the other boiled samples and was selected for purification of anti–SE MAb for detection of boiled SE cells. For LM, the clone of 2F6–7 that reacted strongest with boiled LM 4b was obtained and the culture supernatant was used for purification of anti–LM 4b MAb. The clone of 13H9–2 was found to generate almost greatest response against sonicated LM 1/2a and low response to sonicated L. innocua. Moreover, this MAb reacted strongly with boiled LM 4b without cross–reactivity against the other bacteria. This clone was selected for purification of anti–LM MAb for the detection of sonicated LM 1/2a or LM 4b cells and boiled LM 4b cells. Although MAbs for LM showed cross–reactivity against L. innocua, the MAbs obtained after screening by the combined method showed a capacity to detect target pathogens by using SPR biosensor as well as ELISA. These MAbs are useful for detection of pathogens by biosensor and are expected to contribute to the development of rapid detection of the pathogens.

Original languageEnglish
Pages (from-to)319-330
Number of pages12
JournalJournal of the Faculty of Agriculture, Kyushu University
Volume63
Issue number2
DOIs
Publication statusPublished - 2018

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Agronomy and Crop Science

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