Compositions and methods for protein detection
A protein and mixture technology, applied in the field of mass spectrometry, can solve the problems of inability to distinguish similar transgenic proteins to distinguish transgenic proteins from wild-type proteins, false positives, impossible, etc.
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[0143] Example 1 - Surrogate Peptide Selection
[0144] MRM-based assays rely on the selection of a predetermined set of peptides and depend on specific fragmentation / transition ions for each selected surrogate peptide. Selecting an appropriate surrogate or signature peptide requires several criteria. First, the proteins that make up the targeting protein cassette must be selected. Second, for each target protein, those peptides that exhibit a good mass spectrometric response and uniquely identify that target protein or its specific modifications (ie, post-translational modifications) must be identified. Third, for each mass spectrometrically appropriate peptide, those transition ions that provide the best signal intensity and uniquely distinguish the surrogate peptide from other peptide species present in the sample must be identified. These criteria are critical for performing MRM-based assays.
[0145] Surrogate peptides from seven transgenic proteins, Cry1Ab, eCry3.1Ab,...
example 2
[0158] Example 2 - Assay for detection of transgenic protein in transgenic plant tissue
[0159] The development of sensitive methods for the direct detection of target proteins is highly desirable for quantitative assessment in biological matrices such as tissues of transgenic plants, eg leaf, grain, root and pollen tissues. Multiple reaction monitoring (MRM) mass spectrometry has emerged as a promising platform for quantifying multiple proteins in a given sample by liquid chromatography (LC) coupled with tandem mass spectrometry (MS / MS / ). MRM assays utilize sequence-specific tandem MS fragmentation of proteolytic peptides, thereby providing highly selective and specific measurements of different target proteins. Despite these advances, accurate quantitative measurements of low-abundance proteins or proteins with specific physicochemical properties that affect separation remain challenging.
[0160] MRM determinations are typically performed on triple quadrupole mass spectro...
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