Quantitative analysis of transgenic proteins
AssigneeDow AgroSciences LLC
InventorsTrent James OMAN, Barry W. SCHAFER, Ryan Christopher HILL, Jeffrey R. GILBERT, Guomin SHAN
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Abstract
The invention relates to methods for quantitative multiplex analysis of complex protein samples from plants using mass spectroscopy. In some embodiments, the disclosure concerns methods for maintaining a transgenic plant variety, for example by analyzing generations of a transgenic plant variety for selective and sensitive quantitation of multiplexed transgenic proteins.
Claims23
A high-throughput method of quantitating one or more protein of interest with known amino acid sequence in a plant-based sample, the method comprising:
- (a) extracting proteins from a plant-based sample;
- (b) digesting proteins extracted from step (a) to obtain peptides;
- (c) separating the peptides in a single step;
- (d) determining a plural of signature peptides from the protein of interest with known amino acid sequence;
- (e) measuring the plural of signature peptides using high resolution accurate mass spectrometry (HRAM MS);
- and
- (f) quantitating the protein of interest with known amino acid sequence based on measurements of the signature peptides.
A high-throughput system for quantitating one or more protein of interest with known amino acid sequence in a plant-based sample, the system comprising:
- (a) a high-throughput means for extracting proteins from a plant-based sample;
- (b) a separation module for separating peptides in a single step;
- (c) a selection module for selecting a plural of signature peptides from the protein of interest with known amino acid sequence;
- and
- (d) a high resolution accurate mass spectrometry (HRAM MS) for measuring the plural of signature peptides.
22-29. (canceled)
Dependencies are inferred from the claim text.