公开(公告)号：US09056895B1

公开(公告)日：2015-06-16

申请号：US13886978

申请日：2013-05-03

申请人： WELLSPRING CLINICAL LAB, INC.

发明人： Calvin Wiese

IPC分类号： G01N30/02 ,  C07K1/22 ,  C07K1/20 ,  C07K1/36 ,  G01N30/00 ,  G01N33/566

CPC分类号： C07K1/22 ,  C07K1/145 ,  C07K1/20 ,  C07K1/34 ,  C07K1/36 ,  G01N30/00 ,  G01N30/02 ,  G01N33/566 ,  Y10T436/25375

摘要： The present invention, referred to as Surface-Free Affinity Purification (SFAP), relates to a system and method for purifying low abundance peptides, including digested plasma proteins. SFAP combines the components and processes of Surface-Free Isolation, Specific Competitive Elution, Dual Epitope Isolation and Protective Solvents in the purification of low abundance proteins, all of which may be performed in a single column. This column is a constant volume Diafiltration Column equipped with stirring that can be driven by an HPLC. Digested plasma may be injected into the Diafiltration Column by the HPLC. Immunoglobulins, Specific Competitive Elutants, Protective Solvents are injected in a series of injections that result in the purified peptides bound to the surfaces of a very-low volume hydrophobic reversed phase trap from which they can be easily injected into mass spectrometry detection instruments.

摘要翻译： 称为无表面亲和纯化（SFAP）的本发明涉及用于纯化低丰度肽（包括消化的血浆蛋白）的系统和方法。 SFAP在低丰度蛋白质的纯化中结合了无表面免除隔离，特异性竞争性洗脱，双重表位分离和保护性溶剂的组分和过程，所有这些都可以在单一柱中进行。 该柱是装有可以通过HPLC驱动的搅拌的恒定体积的渗滤柱。 可以通过HPLC将消化的血浆注入渗析柱。 免疫球蛋白，特异性竞争性洗脱剂，保护性溶剂以一系列注射进行注射，其导致纯化的肽结合到非常低体积的疏水性反相捕获物的表面，从而可以将它们轻松地注入质谱检测仪器。