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1. 发明申请

US20100062538A1 IDENTIFYING AND COUNTING PROTEINS IN A SAMPLE 审中-公开
标题翻译：在样品中鉴定和计数蛋白质
公开(公告)号：US20100062538A1
公开(公告)日：2010-03-11
申请号：US12536476
申请日：2009-08-05
申请人： GONGXIN YU , ERIC BROWN , HENRY A. CHARLIER
发明人： GONGXIN YU , ERIC BROWN , HENRY A. CHARLIER
IPC分类号： G01N33/00
CPC分类号： G01N33/6848 , G01N33/6803 , G01N2030/8831
摘要： The proteins in a cell are preferably proteolytically cleaved and chemically attached to another peptide of unique and known sequence. In one embodiment of the invention, peptide-linker-peptide triplets are synthesized with linker molecules such as polyhistidine. In a more preferred embodiment of the invention, peptide-mass differentiated group (MDG) constructs are synthesized. The MDG's may be obtained from a library of oligo-N(K)-peptides synthesized on resin beads, wherein N is the length of the peptides (with a default value of 4) and K is the number of alternative amino acids (with a default value of 10) at each position. Coupling between given peptides and linkers or MDG's creates recombinants with different overall masses that migrate separately in chromatographic separations. The peptides-linker/MGD's recombinants may be purified and sequenced by MS/MS analysis. The resulting purified and sequenced peptides are then counted, and the ratios of the different peptides within and/or between samples obtained.
摘要翻译：细胞中的蛋白质优选蛋白水解切割并化学连接到另一种唯一和已知序列的肽上。在本发明的一个实施方案中，肽 - 接头 - 肽三联体与接头分子如多组氨酸合成。在本发明的更优选的实施方案中，合成肽 - 质量分化组（MDG）构建体。 MDG可以从在树脂珠上合成的寡核苷酸（K）肽文库中获得，其中N是肽的长度（默认值为4），K是替代氨基酸的数目（具有默认值为10）。给定的肽和接头或MDG之间的偶联产生具有不同总质量的重组体，其在色谱分离中单独迁移。肽 - 接头/ MGD的重组体可以通过MS / MS分析进行纯化和测序。然后对所得纯化和测序的肽计数，并且获得的样品内和/或之间的不同肽的比例。

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