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Publication Number
US-8372412-B2
Publication Date
2013-02-12
Expiration Date
Abstract
The invention provides a bioassay for detection of Mycobacterium tuberculosis infection comprising artificially modified peptide subsequences of the T-cell epitope from M. tuberculosis Ag85B. Particularly preferred peptides have the form: SGGNNSPAX (SEQ ID 26), where X is Methionine (SEQ ID 18), Leucine (SEQ ID 17), Alanine (SEQ ID 15) or Valine (SEQ ID 10) and NNSPAV (SEQ ID 14). The invention also provides peptides for use in such an assay.
Core Innovation
The invention relates to peptides derived from the Mycobacterium tuberculosis Ag85B T-cell epitope SEQ ID 11 (GRDIKVQFQSGGNNSPAV) that are optimized for binding to antibodies against that epitope in displacement/competitive ELISA formats. It describes peptide sequences intended for antibody-peptide binding and peptide displacement, including a key 9-mer sequence identified as SGGNNSPAV (SEQ ID 10) that binds tightly.
The disclosed peptides include truncated variants and a peptide library of SGGNNSPAX variants, where SGGNNSPAA (SEQ ID 15), SGGNNSPAL (SEQ ID 17), and SGGNNSPAM (SEQ ID 18) are specifically identified. The content states that GNNSPA (SEQ ID 13) lacks binding, while NNSPAV (SEQ ID 14) binds less tightly than SGGNNSPAV.
The disclosure further identifies improved heat stability for the peptide library variants for field use in hot countries, and it provides assay concepts for detecting M. tuberculosis based on antibody-peptide binding or peptide displacement. Optional peptide labels are described, including fluorescent label examples (e.g., Alexa Fluor), in the context of the antibody-peptide binding/displacement assays.
Claims Coverage
The provided independent claims cover two principal inventive areas: (1) sequence-defined peptide variants consisting of specified short sequences, and (2) antibodies (or antibody fragments) that specifically bind to SEQ ID 11. In total, the independent claims identify two inventive features.
Sequence-defined peptides for antibody binding/displacement
A peptide consisting of SGGNNSPAA (SEQ ID 15), SGGNNSPAL (SEQ ID 17), or SGGNNSPAM (SEQ ID 18).
Antibodies specifically binding to SEQ ID 11
An antibody that specifically binds to SEQ ID 11, or a fragment of said antibody, wherein the said fragment specifically binds to SEQ ID 11.
The claim set (as provided) defines specific short peptide sequences for use in antibody-peptide binding/displacement contexts and also claims antibodies (or fragments) specifically binding to SEQ ID 11.
Stated Advantages
Tightly binding to antibodies against the Ag85B T-cell epitope, as represented by the key 9-mer SGGNNSPAV.
Improved heat stability of peptide library variants for field use in hot countries.
Support for detection of M. tuberculosis using antibody-peptide binding or peptide displacement formats (competitive/displacement ELISA concepts).
Documented Applications
Detecting Mycobacterium tuberculosis using antibody-peptide binding or peptide displacement assay concepts in competitive/displacement ELISA formats.
Assay formats described as antibody-peptide binding/displacement using peptide labels, including optional fluorescent labeling examples.
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