COMPOUNDS AND METHODS FOR PREPARATION OF CONJUGATE REAGENTS

• US 20150072438A1
• Filed: 04/11/2013
• Published: 03/12/2015
• Est. Priority Date: 04/18/2012
• Status: Abandoned Application

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Inventors: Teng, Zhu, Geng, Liping, Drinan, Martin
• Application Number:
  US14/394,634
• Publication Number:
  US 20150072438A1
• Time in Patent Office: Days
• US Class Current: 436/501
• CPC Class Codes: A61K 47/6939 the polymer being a polysaccharide, e.g. starch, chitosan, chitin, cellulose or pectin
• CPC Class Codes: G01N 2333/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin G01N2333/665, corticotropin G01N2333/695
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

Compositions are disclosed that include a support having on a surface thereof at least an inner polysaccharide layer and an outer polysaccharide layer. The outer polysaccharide layer has pendant moieties each comprising a terminal amine functionality. The point of linkage of the pendant moiety to the outer polysaccharide layer does not comprise an amide bond. The composition may be employed in a method of conjugating a member of a specific binding pair to a particle. The method comprises reacting the terminal amine functionality of the composition with the member of the specific binding pair, which comprises an amine-reactive functionality.

Claims

1. A composition comprising a support having on a surface thereof at least an inner polysaccharide layer and an outer polysaccharide layer wherein the outer polysaccharide layer has pendant moieties each comprising a terminal amine group wherein the point of linkage of the pendant group to the outer polysaccharide layer does not comprise an amide bond.

2. The composition according to claim 1 wherein the support is a particle.

3. The composition according to claim 1 wherein the inner polysaccharide layer is an aminodextran and the outer polysaccharide layer is a dextran aldehyde.

4. The composition according to claim 1 wherein the pendant moiety comprises an alkylene or a poly(alkylene ether).

5. A method of conjugating a member of a specific binding pair to a support, the method comprising reacting the terminal amine group of the composition of claim 1 with the member of the specific binding pair that comprises an amine-reactive functionality.

6. The method according to claim 5 wherein the member of the specific binding pair is a hapten.

7. The method of claim 6 wherein the hapten is a hormone.

12. The method according to claim 1 wherein the pendant moiety comprises an alkylene chain or a poly(alkylene ether) chain.

8. A method of forming a conjugate of a hapten and a particle wherein the particle comprises an inner polysaccharide layer and an outer polysaccharide layer that comprises an aldehyde group, the method comprising forming a linkage between the hapten functionalized with an amine-reactive functionality and the aldehyde group with a linking compound comprising two terminal amine groups wherein one amine group of the terminal amine groups reacts with the aldehyde group to form a functionality that does not comprise an amide bond and the other amine group of the terminal amine groups reacts with the amine-reactive functionality.

9. The method according to claim 8, which comprises:

reacting the aldehyde group with one amine group of the terminal amine groups to form a particle with a pendant moiety comprising the other amine group of the terminal amine groups and

reacting the other amine group of the terminal amine groups with the amine-reactive functionality to form the conjugate.

10. The method according to claim 8, which comprises:

reacting one amine group of the terminal amine groups with the amine-reactive functionality to form a hapten having a moiety comprising the other amine group of the terminal amine groups and

reacting the aldehyde group with other amine group of the terminal amine groups to form the conjugate.

11. The method according to claim 8 wherein the inner polysaccharide layer is an aminodextran and the outer polysaccharide layer is a dextran aldehyde.

13. A method of forming a conjugate of a steroid hormone and a particle wherein the particle comprises an inner aminodextran layer and an outer dextran aldehyde layer, the method comprising:

forming a pendant moiety on the particle by reacting an amine group of a linking compound, which comprises an alkylene chain or poly(alkylene ether) chain and terminal amine groups, with an aldehyde group of the outer dextran aldehyde wherein a point of linkage of the pendant moiety to the outer dextran aldehyde layer does not comprise an amide bond linkage and wherein the pendant moiety comprises a terminal amine group, and

reacting the terminal amine group of the pendant moiety with the steroid hormone comprising an amine-reactive functionality to form the conjugate of the steroid hormone and the particle.

14. The method according to claim 13 wherein the particles are latex particles.

15. The method according to claim 13 wherein the particles comprise a label.

16. The method according to claim 15 wherein the label is selected from the group consisting of chemiluminescent compositions and sensitizer compositions.

17. The method according to claim 13 wherein the steroid hormone is selected from the group consisting of progestogens, estrogens, androgens, glucosteroids, mineralocorticoids, and secosteroids.

18. The method according to claim 13 wherein the amine reactive functionality of the steroid hormone is a functionalized ketone, a functionalized aldehyde, a functionalized alcohol, or an epoxy group.

19. The method according to claim 13 wherein the amine reactive functionality of the steroid hormone is carboxymethoxylamine-functionalized.