公开(公告)号：US20070161117A1

公开(公告)日：2007-07-12

申请号：US10587416

申请日：2005-01-26

Applicant: Johan Axelman ,  Anders Ljunglof

Inventor： Johan Axelman ,  Anders Ljunglof

IPC: G01N24/00 ,  G01N1/18

CPC classification number: G01N30/7233 ,  G01N30/02 ,  G01N30/461 ,  G01N30/463 ,  Y10T436/24 ,  Y10T436/255 ,  B01D15/362 ,  B01D15/325 ,  B01D15/363

Abstract: The present invention relates to a method for reducing total sample complexity in native or digested biological sample(s), before analysis thereof by mass spectrometry, comprising the following steps: a) selecting a fraction from the entire native or digested biological sample(s) on the basis of pi-value, said fraction comprising native or digested sample representing a subset of or the entire substance population in the sample; b) separating native or digested sample substances from each other; and c) analysing said substances by mass spectrometry. The invention also relates to a system for reducing total sample complexity in the above method, comprising a high capacity charge-selective column coupled to a MDLC work flow path comprising a cation exchange column and a RPC column. The system is followed by a MS/MS instrument.

Abstract translation: 本发明涉及一种在通过质谱分析之前降低天然或消化的生物样品中的总样品复杂性的方法，包括以下步骤：a）从整个天然或消化的生物样品中选择一个级分， 基于pi值，所述级分包含表示样品中的子集或全部物质群的天然或消化样品; b）将天然或消化的样品物质彼此分离; 和c）通过质谱分析所述物质。 本发明还涉及一种用于降低上述方法中的总样本复杂度的系统，其包括耦合到包括阳离子交换柱和RPC列的MDLC工作流路的高容量电荷选择性列。 该系统后面是MS / MS仪器。

公开(公告)号：US20090242750A1

公开(公告)日：2009-10-01

申请号：US12302381

申请日：2007-06-08

Applicant: Johan Axelman ,  Daniel Ivansson ,  Staffan Renlund

Inventor： Johan Axelman ,  Daniel Ivansson ,  Staffan Renlund

IPC: H01J49/26 ,  B01D59/44

CPC classification number: C07K1/18 ,  C07K1/20 ,  C07K1/36

Abstract: The present invention relates to a method for protein purification and identification. More closely the invention relates to a method for protein pre-fractionation and identification resulting in improved yield of identified proteins. The method for pre-fractionation of protein samples, includes the following steps: a) reducing disulphide bridges (S-S bridges) or protecting cysteines in the proteins in the sample; b) loading the sample onto an ion exchange column; c) eluting the sample; d) collecting each fraction from the column separately in air sealed containers devoid of chromatographic media; e) desalting each fraction on a single RPC (reversed phase chromatography) trap column; f) separating each fraction in a second dimension RPC step to obtain further separated proteins which are collected in fractions; and g) identifying the further separated proteins by MS.

Abstract translation: 本发明涉及一种蛋白质纯化和鉴定方法。 本发明更紧密地涉及蛋白质预分选和鉴定的方法，导致鉴定的蛋白质的产量提高。 蛋白质样品预分离方法包括以下步骤：a）减少样品中蛋白质中的二硫键（S-S桥）或保护半胱氨酸; b）将样品装载到离子交换柱上; c）洗脱样品; d）在没有色谱介质的空气密封容器中分别从塔中收集每个级分; e）在单个RPC（反相色谱）陷阱柱上脱盐每个级分; f）在第二维RPC步骤中分离每个级分以获得分离收集的进一步分离的蛋白质; 和g）通过MS鉴定进一步分离的蛋白质。