Compositions and methods for capturing, isolating, detecting, analyzing and quantifying macromolecules

• US 6,458,599 B1
• Filed: 02/18/2000
• Issued: 10/01/2002
• Est. Priority Date: 02/18/2000
• Status: Expired due to Fees

• Current Assignee: Aspira Biosystems, Inc. (Nanomune Technologies, Inc.)
• Original Assignee: Aspira Biosystems, Inc. (Nanomune Technologies, Inc.)
• Inventors: Huang, Chin-Shiou
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/507,300
• Time in Patent Office: 956 Days
• Field of Search: 436/518, 436/524, 436/531, 436/535, 435/4, 427/2.13
• US Class Current: 436/518
• CPC Class Codes: B01J 20/268 Polymers created by use of a template, e.g. molecularly imprinted polymers
• CPC Class Codes: G01N 2600/00 Assays involving molecular imprinted polymers/polymers created around a molecular template
• CPC Class Codes: G01N 33/544 the carrier being organic
• CPC Class Codes: Y10S 436/804 Radioisotope, e.g. radioimmunoassay
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 977/927 Diagnostic contrast agent

Abstract

A method for making an imprint composition capable of capturing a macromolecule of interest, comprising the steps of (a) solidifying a fluidic matrix material capable of retaining shaped cavities when in a solid or semi-solid form in the presence of a template molecule that corresponds to a portion of the macromolecule, thereby yielding a solid or semi-solid matrix material having template molecules entrapped therein; and (b) removing the template molecules from the solid or semi-solid matrix material, yielding an imprint composition capable of capturing the macromolecule.

Claims

1. A method of making an imprint composition capable of capturing a macromolecule of interest, comprising the steps of:

(a) solidifying a fluidic matrix material capable of retaining shaped cavities when in a solid or semi-solid form in the presence of a template molecule that corresponds to a portion of the macromolecule, thereby yielding a solid or semi-solid matrix material having template molecules entrapped therein; and

(b) removing the template molecules from the solid or semi-solid matrix material, yielding an imprint composition capable of capturing the macromolecule.

2. The method of claim 1 in which the fluidic matrix material is a heat-sensitive composition and the solidifying step comprises lowering the temperature of the heat-sensitive composition from above its melting point to below its melting point.

3. The method of claim 2 in which the heat-sensitive composition is selected from the group consisting of hydrogels, agarose, gelatins and moldable plastics.

4. The method of claim 1 in which the fluidic matrix material is a composition comprising a polymerizable compound and the solidifying step comprises polymerizing the polymerizable compound.

5. The method of claim 4 wherein the polymerizable compound is selected from the group consisting of styrene, methyl methacrylate, 2-hydroxyethyl methacrylate, 2-hydroxyethyl acrylate, methyl acrylate, acrylamide, vinyl ether, vinyl acetate, divinylbenzene, ethylene glycol dimethacrylate, ethylene glycol diacrylate, pentaerythritol dimethacrylate, pentaerythritol diacrylate, N,N′

-methylenebisacrylamide, N,N′

-ethylenebisacrylamide, N,N′

-(1,2-dihydroxyethylene)bis-acrylamide and trimethylolpropane trimethacrylate.

6. The method of claim 1 in which the template molecule is immobilized on a solid support or substrate.

7. The method of claim 1 which the solidifying step is carried out under conditions which are denaturing with respect to the template molecule.

8. The method of claim 1 in which the solidifying step is carried out under conditions which are native with respect to the template molecule.

9. The method of claim 1, wherein the template molecule corresponds to a terminal portion of the macromolecule.

10. The method of claim 1, wherein the macromolecule is a polynucleotide and the template molecule comprises an oligonucleotide.

11. The method of claim 1, wherein the macromolecule is a polypeptide and the template molecule comprises an oligosaccharide or a glycopeptide.

12. The method of claim 1, wherein the macromolecule is a polypeptide and the template molecule comprises a peptide.

13. The method of claim 12, wherein the sequence of the peptide corresponds to a contiguous sequence of the polypeptide.

14. The method of claim 13, wherein the peptide is between 3 and 30 amino acids in length.

15. The method of claim 13, wherein the peptide is between 4 and 15 amino acids in length.

16. The method of claim 13, wherein the peptide is between 4 and 7 amino acids in length.

17. The method of claim 13, wherein the portion of the polypeptide comprises the C-terminus of the polypeptide, the N-terminus of the polypeptide or an internal sequence of the polypeptide.