Method of producing probe arrays for biological materials using fine particles

Abstract

The use of probe arrays in which probes of various biological substances such as DNA are immobilized on the surface of a solid is becoming established as an effective means for high-speed screening. Different kinds of probes, such as DNA, are immobilized on the surface of a multiple number of independently treatable fine particles, such as beads, instead of the surface of a single solid, and the resulting beads are aligned in a capillary or a cell in a designated order. The size of the area where one probe is immobilized is reduced. The bead probe array is characterized in that such small beads are aligned one by one in a designated manner using a sheet with holes, and one or a multiple number of beads are held in the holes and then transferred to a probe array holder such as a capillary.

1 . A method for producing a probe array, comprising the steps of: 
 selecting plural types of probes of interest;    immobilizing the plural types of probes on surfaces of different solid pieces; and    aligning the probe-immobilized solid pieces in a designated order to obtain a probe array for analyzing a sample solution passing therethrough, wherein said aligning is performed by the following steps:    (i) placing the probe-immobilized solid pieces in a well on a sheet having a hole through which one solid piece can pass, said hole leading to the interior of the array, said hole being closed;    (ii) trapping one of the solid pieces in said hole:    (iii) opening the hole;    (iv) transferring the trapped solid piece to the array; and    (v) repeating steps (i) through (iv) until a designated number of arrays are filled with the probe-immobilized solid pieces aligned therein.    
     
     
         2 . The method according to  claim 1 , wherein the probes are polynucleotides, peptides, or proteins.  
     
     
         3 . The method according to  claim 1 , wherein the solid pieces are beads.  
     
     
         4 . The method according to  claim 3 , wherein the beads are fine particles.  
     
     
         5 . The method according to  claim 1 , wherein the alignment of the solid pieces is a one-dimensional arrangement or a two-dimensional arrangement.  
     
     
         6 . The method according to  claim 1 , wherein the alignment is conducted in an array selected from the group consisting of a capillary, a groove, and an optical cell.  
     
     
         7 . The method according to  claim 1 , wherein said sheet is placed on a movable base having a through-hole leading into the array holder, said movable base being positioned where the hole of the sheet does not communicate with the through-hole of the movable base; said method further comprising after step (ii) 
 removing the remaining solid pieces from the sheet; and    moving the movable base to a position where the hole of the sheet communicates with the through-hole of the movable base.    
     
     
         8 . The method according to  claim 1 , wherein said hole is closed with a valve and wherein opening said hole is performed by opening said valve.  
     
     
         9 . The method according to  claim 1 , wherein each well contains a single type of probe-immobilized solid pieces, each well having a hole through which one solid piece can pass, said hole being clos, said method further comprising 
 closing each hole after moving the wells in a designated order to transfer each trapped solid piece to an array;    moving the wells to align the probe-immobilized solid pieces in a next array; and    repeating all steps through until a designated number of arrays are filled with the probe-immobilized solid pieces aligned therein.