Method and device for investigating substance libraries

• US 7,431,685 B2
• Filed: 03/28/2001
• Issued: 10/07/2008
• Est. Priority Date: 03/28/2000
• Status: Expired due to Fees

• Current Assignee: Borstel Forschungszentrum
• Original Assignee: Laser-Und Medizin-Technologie Gmbh Berlin
• Inventors: Mueller, Gerhard, Frey, Andreas, Helfman, Juergen, Schmidt, Marcus Alexander
• Primary Examiner(s): Schultz; J D
• Assistant Examiner(s): Lundgren; Jeffrey S.
• Application Number:
  US10/239,986
• Publication Number:
  US 20030165998A1
• Time in Patent Office: 2,750 Days
• Field of Search: 435/4, 435/7.1, 435/287.3, 435/287.9, 435/288.2, 435/288.3, 435/DIG.46, 435/DIG.49
• US Class Current: 506/23
• 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 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00306 Reactor vessels in a multiple arrangement
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00353 Pumps
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00414 using suction
• CPC Class Codes: B01J 2219/00418 using pressure
• CPC Class Codes: B01J 2219/00486 by sonication or ultrasonication
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/0052 in the shape of elongated tubes
• CPC Class Codes: B01J 2219/00522 in a multiple parallel arrangement
• CPC Class Codes: B01J 2219/00527 Sheets
• 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 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00621 : by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 : Covalent
• CPC Class Codes: B01J 2219/00637 : by coating it with another layer
• CPC Class Codes: B01J 2219/00659 : Two-dimensional arrays
• CPC Class Codes: B01J 2219/00707 : separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: C07B 2200/11 : Compounds covalently bound to a solid support
• CPC Class Codes: C07K 1/045 : using devices to improve synthesis, e.g. reactors, special vessels
• CPC Class Codes: C07K 1/047 : Simultaneous synthesis of different peptide species; Peptide libraries
• 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 60/14 : for creating libraries

Abstract

The aim of the invention is to investigate the bonding of substances to target molecules. This is achieved by means of a densely packed device wherein various target molecules are bonded in a large number of sample areas. Cross-contamination and evaporation need to be minimized. The active surface of the sample areas have to be maximized. The inventive solution resides in the use of carrier plates, containing densely packed capillary structures and having a very large inner surface despite small outer dimensions. 1000 times more molecules can be bonded than on a flat outer surface of a comparable size. Cross contamination is avoided by the lack of cross links between the capillaries. Evaporation is minimized by a small outer surface. After the inner surface of the capillaries has been silanized, peptides and peptidomimetics are synthesized in a locally targeted manner. The molecular interactions of components of the substance library with active substances in a solution or a suspension are investigated by means of a local resolution optical detection method. Handling, especially cleaning and covering with substances, is carried out in a simple manner by rinsing liquids through the capillary plate.

Claims

1. A method of preparing a substance library for investigating the molecular interaction of soluble or suspendable substances with solid-phase bounded peptidic or peptoid target molecules wherein the substance library comprises a plurality of parallel capillaries arranged in a plate, the method comprising the following steps:

a) applying an organosilane layer on an internal surface of the capillaries, said organosilane layer having functional groups for anchoring peptidic or peptoid target molecules;

b) defining sample areas wherein each sample areas comprises a plurality of adjacent capillaries, and wherein defining said sample areas is performed by position dependent application of a protecting substance temporarily protecting the functional groups of the organosilane layer of the capillaries within said sample areas or by position-dependent application of a protected anchor molecule within said sample areas;

c) saturating the capillaries in the plate outside the sample areas with a deactivation reagent such that neither synthesis nor coupling of target molecules can occur in the capillaries outside the sample areas;

d) deprotecting all protected functional groups or anchor molecules in the sample areas; and

e) synthesizing peptidic or peptoid target molecules in the sample areas by pipetting solutions into the capillaries of the sample areas;

wherein said solutions contain amino acids for sequential synthesis of peptide or peptoid target molecules or wherein said solutions are coupling solutions for coupling complete peptide or peptoid target molecules.

2. The method of claim 1, wherein a protected α

-aminopoly(ethyleneglycol)-ω

-carboxylic acid active ester is used as the protected anchor molecule in step b).

3. The method of claim 2, wherein fluorenylmethoxycarbonyl (FMOC)-protected α

-aminopoly(ethyleneglycol)-ω

-propionic acid-N-hydroxybenzotriazole ester is used as the protected anchor molecule in step b).

4. The method of claim 1, wherein the capillary plate comprises metal or semi-metal, and wherein prior to applying an organosilane layer according to step a), an oxide layer is provided for binding the silane by means of surface oxidation of the internal surface of the capillaries.

5. The method of claim 1, wherein a plurality of sample areas are each of a diameter of up to 2,000 μ

m.

6. The method of claim 1, wherein the capillaries are of a length and the capillary plate is of a thickness of between 100 and 2,000 μ

m.

7. The method of claim 1, wherein up to 4,000 adjacent capillaries are present in a sample area.