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Development of a novel engineered antibody targeting Neisseria species

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Abstract

Objectives

A Neissaria bacterial pilus sugar, bacillosamine, was synthesized and, for the first time, used as a probe to screen a single-chain variable fragment (scFv).

Results

Four Neisseria, Neisseria gonorrhoeae, Neisseria meningitidis, Neisseria sicca and Neisseria subflava, and two negative controls, Streptococcus pneumoniae and Escherichia coli, were tested through ELISA, immunostaining and gold nanoparticle immunological assay. All results indicated that the selected scFv is feasible for the specific detection of Neisseria species via the recognition of bacillosamine.

Conclusions

The recombinant scFv could detect Neisseria strains at 106 CFU/ml.

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Acknowledgements

This work was financially supported by Grants from the Ministry of Science and Technology, Taiwan (MOST 105-2113-M-009-005), and the Center for interdisciplinary science, National Chiao Tung University.

Supporting information

Supplementary Methods—Compound 1 synthesis and BSA modification. Screening of compound 1-specific clones by monoclonal phage ELISA. Bacterial strain and culture conditions. Isolation of Neisseria pili. Characterization of purified recombinant anti-compound 1 scFv antibody. Immunofluorescence staining and confocal laser scanning microscopy. Nano-gold particle immunological assay. Supplementary Scheme 1—Reagents and conditions of compound 1 synthesis. Supplementary Scheme 2—Installation of compound 1 to BSA. Supplementary Fig. 1—Procedure used to select compound 1-specific recombinant scFv antibodies. Supplementary Fig. 2—Approach used to determine scFv binding activity to compound 1. Supplementary Fig. 3—Immunofluorescence of Neisseria species. (Incubation with fluorescein were considered as the background.)

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Authors and Affiliations

  1. Department of Applied Chemistry, National Chiao Tung University, Hsinchu, 30010, Taiwan

    Chih-Yueh Liu, Chang-Ching Weng, Chih-Hsiang Lin, Kwok-Kong Tony Mong & Yaw-Kuen Li

  2. Institute of Molecular Biology, National Chung Hsing University, Taichung, 40227, Taiwan

    Chiou-Ying Yang

Authors
  1. Chih-Yueh Liu
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  2. Chang-Ching Weng
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  3. Chih-Hsiang Lin
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  4. Chiou-Ying Yang
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  5. Kwok-Kong Tony Mong
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  6. Yaw-Kuen Li
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Correspondence to Yaw-Kuen Li.

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Cite this article

Liu, CY., Weng, CC., Lin, CH. et al. Development of a novel engineered antibody targeting Neisseria species. Biotechnol Lett 39, 407–413 (2017). https://doi.org/10.1007/s10529-016-2258-1

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  • DOI: https://doi.org/10.1007/s10529-016-2258-1

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