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Real time detection of anthrax spores using highly specific anti-EA1 recombinant antibodies produced by competitive panning.
J Immunol Methods 2008; 334(1-2):1-10JI

Abstract

We describe a targeted approach for the production of biological recognition elements capable of fast, specific detection of anthrax spores on biosensor surfaces. The aim was to produce single chain antibodies (scFvs) to EA1, a Bacillus anthracis S-layer protein that is also present, although not identical, in related to Bacillus species. The aim of the work was to produce antibodies that would detect B. anthracis EA1 protein and intact spores with a high degree of specificity, but would not detect other Bacillus species. Existing monoclonal antibodies were evaluated and found to recognise B. anthracis EA1 and S-layer proteins from other closely related Bacillus species. Recombinant anti-EA1 scFvs were isolated from B. anthracis immune library that contained antibody genes raised against B. anthracis spores and purified exosporium. Two approaches for scFv selection were used; standard (non-competitive) panning, and competitive panning. The non-competitive biopanning strategy isolated scFvs that recognised EA1 from B. anthracis, but also cross-reacted with other Bacillus species. In contrast, the competitive panning approach used S-layer proteins from other Bacillus species to generate scFvs that were highly specific to B. anthracis EA1 and demonstrated apparent nanomolar binding affinities. Specific, real time detection of B. anthracis spores was demonstrated with these scFvs using an evanescent wave biosensor, the Resonant Mirror. The approach described can be used to generate specific antibodies to any desired target where homologous proteins also exist in closely related species, and demonstrates clear advantages to using recombinant technology to produce biological recognition elements for detection of biological threat agents.

Authors+Show Affiliations

Detection Department, Dstl Porton Down, Salisbury, Wiltshire, SP4 OJQ, United Kingdom. telove@dstl.gov.ukNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18395220

Citation

Love, Tracey E., et al. "Real Time Detection of Anthrax Spores Using Highly Specific anti-EA1 Recombinant Antibodies Produced By Competitive Panning." Journal of Immunological Methods, vol. 334, no. 1-2, 2008, pp. 1-10.
Love TE, Redmond C, Mayers CN. Real time detection of anthrax spores using highly specific anti-EA1 recombinant antibodies produced by competitive panning. J Immunol Methods. 2008;334(1-2):1-10.
Love, T. E., Redmond, C., & Mayers, C. N. (2008). Real time detection of anthrax spores using highly specific anti-EA1 recombinant antibodies produced by competitive panning. Journal of Immunological Methods, 334(1-2), pp. 1-10. doi:10.1016/j.jim.2007.12.022.
Love TE, Redmond C, Mayers CN. Real Time Detection of Anthrax Spores Using Highly Specific anti-EA1 Recombinant Antibodies Produced By Competitive Panning. J Immunol Methods. 2008 May 20;334(1-2):1-10. PubMed PMID: 18395220.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Real time detection of anthrax spores using highly specific anti-EA1 recombinant antibodies produced by competitive panning. AU - Love,Tracey E, AU - Redmond,Caroline, AU - Mayers,Carl N, Y1 - 2008/02/29/ PY - 2005/10/10/received PY - 2006/03/01/accepted PY - 2008/4/9/pubmed PY - 2008/7/9/medline PY - 2008/4/9/entrez SP - 1 EP - 10 JF - Journal of immunological methods JO - J. Immunol. Methods VL - 334 IS - 1-2 N2 - We describe a targeted approach for the production of biological recognition elements capable of fast, specific detection of anthrax spores on biosensor surfaces. The aim was to produce single chain antibodies (scFvs) to EA1, a Bacillus anthracis S-layer protein that is also present, although not identical, in related to Bacillus species. The aim of the work was to produce antibodies that would detect B. anthracis EA1 protein and intact spores with a high degree of specificity, but would not detect other Bacillus species. Existing monoclonal antibodies were evaluated and found to recognise B. anthracis EA1 and S-layer proteins from other closely related Bacillus species. Recombinant anti-EA1 scFvs were isolated from B. anthracis immune library that contained antibody genes raised against B. anthracis spores and purified exosporium. Two approaches for scFv selection were used; standard (non-competitive) panning, and competitive panning. The non-competitive biopanning strategy isolated scFvs that recognised EA1 from B. anthracis, but also cross-reacted with other Bacillus species. In contrast, the competitive panning approach used S-layer proteins from other Bacillus species to generate scFvs that were highly specific to B. anthracis EA1 and demonstrated apparent nanomolar binding affinities. Specific, real time detection of B. anthracis spores was demonstrated with these scFvs using an evanescent wave biosensor, the Resonant Mirror. The approach described can be used to generate specific antibodies to any desired target where homologous proteins also exist in closely related species, and demonstrates clear advantages to using recombinant technology to produce biological recognition elements for detection of biological threat agents. SN - 0022-1759 UR - https://www.unboundmedicine.com/medline/citation/18395220/Real_time_detection_of_anthrax_spores_using_highly_specific_anti_EA1_recombinant_antibodies_produced_by_competitive_panning_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-1759(08)00051-3 DB - PRIME DP - Unbound Medicine ER -