QUANTITATIVE ANALYSIS OF TRANSGENIC PROTEIN 5-ENOLPYRUVYLSHIKIMATE-3-PHOSPHATE SYNTHASE

摘要

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.

主权项

1. 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.