Polyallylamine conjugates and applications thereof for biological signal amplification

• US 7,811,571 B2
• Filed: 10/15/2004
• Issued: 10/12/2010
• Est. Priority Date: 11/13/2003
• Status: Expired due to Fees

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research Institute
• Inventors: Pan, Chung We, Tsao, Jia Huey, Yin, Li Te, Tsao, Chao Yun, Chiou, Su Fung
• Primary Examiner(s): Noakes; Suzanne M
• Assistant Examiner(s): Lee; Jae W
• Application Number:
  US10/964,713
• Publication Number:
  US 20050106649A1
• Time in Patent Office: 2,188 Days
• Field of Search: None
• US Class Current: 424/178.1
• CPC Class Codes: G01N 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding

Abstract

Polyallylamine conjugates and applications thereof for biological signal amplification are provided by utilizing the essential amino group of polyallylamine to covalently bind with capture agents and signal molecules having the functional groups selected from a group consisting of —NHS, —CO, —S═O₂ and —C═O—C═O. The resulting conjugates having more than one signaling entities can be further implemented for biological expression with enhancing effect on biological signal intensity, such that the sensitivity of detection for the variation between biological interactions is largely increased.

Claims

1. A polyallylamine conjugate, consisting of polyallylamine, a signal molecule, and a capture agent, wherein said signal molecule is a fluorescent molecule which is cy5;

said capture agent is a modified antibody linked to a cross-linking agent, wherein the antibody is modified by a compound having functional groups selected from the group consisting of —

SH, —

CO, —

NH₂ and —

N═

C═

S, and a cross-linking agent selected from the group consisting of N-succinimidyl 3-(2-pyridyldithio) propionate, glutaraldehyde, succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), maleimide, iodoacetamide, protected thiol and sulfosuccinimidyl-6-(3′

-[2-pyridyldithio] propionamido) hexanoate (sulfo-LC-SPDP), and wherein said cross-linking agent assists the binding of said capture agent with the —

NH₂ functional groups of polyallylamine; and

the signal molecule and capture agent is covalently bound to the polyallylamine via —

NH₂ side chain of the polyallylamine.

3. The polyallylamine conjugate according to claim 1, wherein the polyallylamine conjugate is in contact with a target molecule.

4. The polyallylamine conjugate according to claim 3, wherein the target molecule is an antigen or on a biochip.

5. The polyallylamine conjugate according to claim 4, wherein the target molecule is an antigen.

7. The polyallylamine conjugate according to claim 3, wherein the antibody is an anti-IgE antibody.

8. The polyallylamine conjugate according to claim 4, wherein the antibody is an anti-IgE antibody.

9. The polyallylamine conjugate according to claim 5, wherein the antibody is an anti-IgE antibody.

2. A polyallylamine conjugate, consisting essentially of polyallylamine, a signal molecule, and a capture agent, wherein said signal molecule is a fluorescent molecule which is cy5;

said capture agent is a modified antibody linked to a cross-linking agent, wherein the antibody is modified by a compound having functional groups selected from the group consisting of —

SH, —

CO, —

NH₂ and —

N═

C═

S, and a cross-linking agent selected from the group consisting of N-succinimidyl 3-(2-pyridyldithio) propionate, glutaraldehyde, succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), maleimide, iodoacetamide, protected thiol and sulfosuccinimidyl-6-(3′

-[2-pyridyldithio] propionamido) hexanoate (sulfo-LC-SPDP), and wherein said cross-linking agent assists the binding of said capture agent with the —

NH₂ functional groups of polyallylamine; and

the signal molecule and capture agent is covalently bound to the polyallylamine via —

NH₂ side chain of the polyallylamine.

6. The polyallylamine conjugate according to claim 2, wherein the antibody is an anti-IgE antibody.

10. The polyallylamine conjugate according to claim 2, wherein the polyallylamine conjugate is in contact with a target molecule.

11. The polyallylamine conjugate according to claim 10, wherein the target molecule is an antigen or on a biochip.

12. The polyallylamine conjugate according to claim 11, wherein the target molecule is an antigen.

13. The polyallylamine conjugate according to claim 12, wherein the antibody is an anti-IgE antibody.

14. The polyallylamine conjugate according to claim 13, wherein the antibody is an anti-IgE antibody.

15. The polyallylamine conjugate according to claim 14, wherein the antibody is an anti-IgE antibody.