Pipetting device with pipette tip for solid phase reactions

• US 6,143,252 A
• Filed: 04/12/1999
• Issued: 11/07/2000
• Est. Priority Date: 04/12/1999
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Perkinelmer Incorporated
• Inventors: Hoe, Liang Boon, Haxo, Francis Theodore Jr.
• Primary Examiner(s): Howard, Jacqueline V.
• Assistant Examiner(s): OHORODNIK, SUSAN K
• Application Number:
  US09/292,777
• Time in Patent Office: 575 Days
• Field of Search: 73/863.31-863.33, 73/864.87, 73/864.16, 73/864.17, 422/63, 422/100, 422/135, 422/134, 422/131, 422/923, 436/180, 128/DIG. 1
• US Class Current: 506/40
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/0029 with pistons or plungers
• CPC Class Codes: B01J 2219/00313 the reactor vessels being formed by arrays of wells in blocks
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00353 Pumps
• CPC Class Codes: B01J 2219/00369 in multiple or parallel arrangements
• CPC Class Codes: B01J 2219/00414 using suction
• CPC Class Codes: B01J 2219/00418 using pressure
• CPC Class Codes: B01J 2219/00423 using filtration, e.g. through porous frits
• CPC Class Codes: B01J 2219/00454 by chemical cleavage from the solid support
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00686 Automatic
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 3/0206 : of the plunger pump type
• CPC Class Codes: B01L 3/0217 : of the plunger pump type medical syringes A61M
• CPC Class Codes: B01L 3/0279 : co-operating with positive ejection means
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2035/1055 : for immobilising reagents, e.g. dried reagents

Abstract

A pipetting device for use with a pipette tip containing a solid phase reaction support, comprising a cylinder, a plunger movable within the cylinder between a retracted and an extended position, and a seal disposed between the plunger and cylinder for holding the plunger as it moves. The plunger has a central passageway within an upper portion of the plunger communicating with a side egress port on the plunger, the plunger movable between a retracted position, for aspirating and dispensing liquid reagent wherein the port remains above the seal, and an inserted position wherein the port is positioned below the seal for allowing fluid (e.g., washing solution or inert gas) to be introduced into the cylinder and tip. In an apparatus and method, an array of such devices is used, each fitted with a pipette tip having a solid support retained therein, for performing high-throughput and parallel chemical synthesis of biopolymers (e.g., oligonucleotides or peptides). In one aspect, means are provided for separately controlling the movement of each device in the apparatus for contacting liquid reagent with the solid support. In another aspect, a tip remover is provided for selectively removing tips containing completed polymer.

Claims

1. A pipetting device for solid phase synthesis, said device comprising

a cylinder having an upper end and a lower end and defining a cylindrical chamber extending therebetween,

a plunger having an upper end and a lower end, wherein the plunger (i) is slidably movable in the cylindrical chamber between a retracted and an inserted position, (ii) has an outer diameter that is less than the inner diameter of the chamber, and wherein the difference between the outer diameter of the plunger and the inner diameter of the cylindrical chamber is large enough to allow fluid passage and (iii) has a central passageway leading from a portion of the plunger above said chamber to an egress port which traverses a lateral wall of the plunger within said chamber,

a seal disposed adjacent the upper end of said cylinder for holding said plunger during movement thereof, and

a pipetting terminus for mounting a disposable pipette tip, wherein said plunger is movable between (i) a retracted position wherein said egress port is positioned above said chamber, and (ii) an inserted position wherein said egress port is positioned within said chamber, for introducing a fluid through said passageway into the cylindrical chamber for delivery to a pipette tip when a pipette tip is mounted on said pipetting terminus.

2. The device of claim 1, which further includes a pipette tip mounted on said pipetting terminus, and said pipette tip contains a solid support for solid phase synthesis.

3. The device of claim 2, wherein said solid support contains at least one blocked protected nucleoside attached covalently thereto.

4. The device of claim 2, wherein said solid support contains at least one protected amino acid residue attached covalently thereto.

5. An apparatus for solid phase synthesis, said apparatus comprising:

a plurality of devices in accordance with claim 1, wherein the plungers of said devices are operatively attached to a plunger plate for simultaneously moving the plungers relative to said cylindrical chambers for liquid dispensing.

6. An apparatus for solid phase synthesis, said apparatus comprising:

a plurality of devices in accordance with claim 2, wherein the plungers of said devices are operatively attached to a plunger plate for simultaneously moving the plungers relative to said cylindrical chambers for liquid dispensing.

7. An apparatus for solid phase synthesis, said apparatus comprising:

a plurality of devices in accordance with claim 1, wherein the movement of each device is under separate control of a controller so that each said devices can be manipulated independently of the other devices in the apparatus.

8. The apparatus of claim 7, which further includes:

a power plate for moving the cylinder(s) of one or more of said devices longitudinally, said plate further defining apertures through which the cylinders protrude, and

locking means associated with each cylinder for reversibly and separately locking each cylinder to said power plate.

9. The apparatus of claim 8, wherein each cylinder includes one or more pin members for locking with the power plate, the power plate includes corresponding recesses for allowing said pin members to traverse the power plate for each cylinder that is in an unlocked position, and the apparatus further includes means for independently rotating each cylinder about its longitudinal axis so that said pin member(s) may be locked to or unlocked from the power plate.

10. The apparatus of claim 7, which further includes a plurality of drive motors each having a drive shaft connected to anassociated plunger for independent longitudinal movement of each plunger.