Covalent bonding of molecules to an activated solid phase material

• US 6,306,665 B1
• Filed: 10/13/1999
• Issued: 10/23/2001
• Est. Priority Date: 10/13/1999
• Status: Expired due to Term

• Current Assignee: Quantrx Biomedical Corporation
• Original Assignee: A-Fem Medical Corporation
• Inventors: Hyatt, Timothy Patrick, Buck, Robert Lee, Mueggler, Paul Andrew, Wang, Huiying
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): GABEL, GAILENE
• Application Number:
  US09/417,957
• Time in Patent Office: 741 Days
• Field of Search: 422/55, 422/56, 422/57, 422/58, 422/60, 422/61, 422/62, 422/68.1, 422/69, 435/4, 435/6, 435/7.1, 435/7.5, 435/7.92, 435/128, 435/130, 435/286.5, 435/287.2, 435/287.3, 435/287.7, 435/287.8, 435/287.9, 436/518, 436/524, 436/527, 436/528, 436/529, 436/530, 436/532, 436/535, 436/538, 436/541, 436/162, 436/166, 436/168, 436/169, 436/170, 436/808, 559/419, 559/421
• US Class Current: 436/530
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387

Abstract

Microporous solid phase materials that are suitable for lateral flow and other assays for detecting the presence of analytes in test samples, that are stable under variations in humidity and, even after storage for extended periods of time, can form stable covalent bonds with molecules containing a free primary or secondary amine group or sulfhydryl group are described. The invention further concerns chemically derivatized solid phase materials, and conjugates comprising such materials. Examples of lateral flow devices for the quantitative or semi-quantitative determination of an analyte in a biological sample are described.

Claims

1. A conjugate having the formula

where X is a hydroxyl bearing solid phase material;

R₅ is (CH₂)_n(NH(CH₂)_m)_p, where n is from 2 to 8, m is from 2 to 8, and p is from 0 to 3;

R₆ is selected from the group consisting of alkyl, cyclic alkyl, aromatic and heterocyclic groups containing 0 to 8 hydroxyl, hydroxycarbonyl, or aminocarbonyl groups; and

Q is any molecule that contains a free primary or secondary amine group.

2. The conjugate of claim 1 where X is selected from the group of nitrocellulose, cellulose, glass fibers and porous glass beads.

3. The conjugate of claim 2 where X is nitrocellulose.

4. The conjugate of claim 1 where R5is a propyl group.

5. The conjugate of claim 1 where R6is an alkyl group.

6. The conjugate of claim 5 where R6is an ethyl group.

7. The conjugate of claim 1 where Q is selected from the group of molecules that intrinsically contain a free primary or secondary amine group and molecules to which a free primary or secondary amine group has been covalently attached.

8. The conjugate of claim 7 where Q intrinsically contains a free primary amine group.

9. A method for preparing the conjugate of claim 1, comprising:

applying Q in an aqueous solution to a chemically derivatized solid phase material having the formula

 

where X is a hydroxyl bearing solid phase material;

R5is (CH2)n(NH(CH2)m)p, where n is from 2 to 8, m is from 2 to 8, and p is from 0 to 3;

R6is selected from the group consisting of alkyl, cyclic alkyl, aromatic and heterocyclic groups containing 0 to 8 hydroxyl, hydroxycarbonyl, or aminocarbonyl groups; and

Y and Z are independently hydrogen, alkyl alkyl carbonyl, aromatic carbonyl or heterocyclic carbonyl groups;

or N, Y, Z together are succinimidyl, benzotriazolyl, or derivatives thereof;

drying the solid phase material;

washing the dried solid phase material; and

redrying the washed solid phase material.

10. The method of claim 9 where X is selected from the group of nitrocellulose, cellulose, glass fibers and porous glass beads.

11. The method of claim 10 where X is nitrocellulose.

12. The method of claim 9 where R5is a propyl group.

13. The method of claim 9 where R6is an alkyl group.

14. The method of claim 13 where R6is an ethyl group.

15. The method of claim 9 where N, Y and Z together are succinimide.

16. The method of claim 9 where N, Y, and Z together are N-hydroxybenzotriazole.