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.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Love TE,Redmond C,Mayers CNdoi
10.1016/j.jim.2007.12.022subject
Has Abstractpub_date
2008-05-20 00:00:00pages
1-10issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(08)00051-3journal_volume
334pub_type
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00168-8
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00103-9
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
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pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
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abstract：:A simple immunoenzyme technique is described in which the enzyme beta-galactosidase of E. coli (Z) is used as a marker. The principle of the test is based on the ability of the cells to bind the antigen. Conditions of the assay using Z both as the antigen and marker are described. As a screening technique, conditions ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
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更新日期：2002-12-15 00:00:00
