Anti-clogging device and method for in-gel digestion applications

Abstract

An integrated proteomics sample preparation device and method for in-gel digestion of proteins and for desalting and concentrating samples prior to further analysis such as by MALDI TOF and/or electro-spray ionization (ESI) mass spectrometry. The device and method of the present invention allow for digestion, desalting and concentration of sample prior to analysis. More specifically, the device in accordance with an embodiment of the present invention includes a plurality of wells in fluid communication with a an outlet or drainage opening containing a three dimensional structure comprising a plurality of sorptive particles entrapped in a porous polymer matrix so as to form a device capable of carrying out solid phase extraction. In a preferred embodiment, the wells are configured so as to prevent a sample carrier present in the wells from clogging the outlet when subjected to a driving force such as vacuum. The device also reduces or eliminates overflowing of a well in the event a drain becomes clogged during automated operation.

1 . A method for sample preparation of sample(s) containing protein for further analysis, comprising: 
 providing a carrier containing said sample;    providing at least one well having an inlet, a bottom surface, a drain in said bottom surface, and at least one passageway formed in said bottom surface about said drain and having a largest dimension smaller than the smallest dimension of said carrier;    supporting said carrier at said bottom surface while retaining fluid communication between said bottom surface and said drain via said at least one passageway;    digesting said protein in said sample in said carrier by adding enzyme solution,    extracting peptides from said carrier by adding a solution to said well;    applying vacuum to said well; and    causing said extracted peptides to flow to said drain through said at least one passageway.    
   
   
       2 . The method of  claim 1 , wherein there are three passageways arranged symmetrically about said drain.  
   
   
       3 . The method of  claim 1 , wherein said peptides are absorbed in said drain by an adsorptive structure positioned in said drain.  
   
   
       4 . The method of  claim 3 , wherein said structure comprises a plurality of sorptive particles entrapped in a porous matrix.  
   
   
       5 . The method of  claim 1 , wherein said carrier comprises a gel.  
   
   
       6 . The method of  claim 1 , wherein said carrier comprises a membrane.  
   
   
       7 . A method for sample preparation of sample containing protein for further analysis, comprising: 
 providing a plurality of carriers containing said sample;    providing an array of wells, each well having an inlet, a bottom surface, a drain in said bottom surface, and at least one passageway formed in said bottom surface about said drain and having a dimension smaller than the smallest dimension of one of said plurality of carriers;    supporting a respective one of said plurality of carriers at said bottom surface of each said well while retaining fluid communication between said bottom surface and said drain via said at least one passageway;    digesting said protein in said sample in each said carrier by adding enzyme solution;    extracting peptides from each said carrier by adding solution to each said well;    simultaneously applying vacuum to said each well; and    causing said eluted peptides to flow from each said carrier to said drain through said at least one passageway in each said well.    
   
   
       8 . The method of  claim 7 , wherein each said carrier comprises a gel.