Polyether polyamines as linking agents for particle reagents useful in immunoassays

• US 4,581,337 A
• Filed: 07/07/1983
• Issued: 04/08/1986
• Est. Priority Date: 07/07/1983
• Status: Expired due to Term

• Current Assignee: Dade Behring, Inc. (Siemens AG)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Simons, Donald M., Frey, William A.
• Primary Examiner(s): NOT, DEFINED
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US06/511,751
• Time in Patent Office: 1,006 Days
• Field of Search: 436/531, 436/532, 436/533, 436/534, 436/517, 436/543, 436/547, 436/805, 436/823, 435/180, 435/181, 427/2, 428/402, 428/403, 260/112 R
• US Class Current: 436/533
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/823 Immunogenic carrier or carrier per se

Abstract

Particle reagents, having polyether polyamine linking groups, for use in turbidimetric immunoassays are provided. These linking agents permit ready covalent attachment of compounds of biological interest to polymer particles and lead to immunoassays of improved precision.

Claims

1. A particle reagent consisting essentially of:

(A) a polymer particle having an inner core and an outer shell wherein the inner core is a polymer having a refractive index of not less than 1.54 as measured at the wavelength of the sodium D line and wherein the outer shell is a polymer of

(1) an ethylenically unsaturated monomer having a functional group selected from the group consisting of epoxy, carboxy, and aldehyde, said functional group capable of reacting with a hydrophilic polymeric linker; and

(2) not more than 10 parts by weight of the outer shell of the monomers of the inner core, said outer shell being formed by polymerization in the presence of said inner core; and

wherein said polymer particle has an approximate diameter range of 0.03-0.1 μ

m and is covalently attached to

(B) a compound of biological interest through a hydrophilic polymeric linker of empirical formula;

space="preserve" listing-type="equation">H[NHC.sub.2 H.sub.3 R(OC.sub.2 H.sub.3 R).sub.x OC.sub.2 H.sub.3 R].sub.y NH.sub.2

wherein R is at least one member selected from the group consisting of H and CH₃, x is 0-70 and y is 1-20.

2. The particle reagent of claim 1 wherein said outer shell additionally contains other ethylenically unsaturated monomers in an amount sufficient to render the polymer particles substantially water insoluble.

3. The particle reagent of claim 1 or claim 2 wherein R═

H.

4. The particle reagent of claim 3 wherein X=1-15 and y=1-15.

5. The particle reagent of claim 4 wherein X=1-2 and y=1-5.

6. The particle reagent of claim 1 or claim 2 wherein the linker is derived from propylene oxide-capped polyethylene glycol.

7. The particle reagent of claim 1 or claim 2 wherein X=1-15 and y=1-15.

8. A method for measuring compounds of biological interest comprising the steps of:

(A) incubating

(1) a particle reagent having high refractive index consisting essentially of;

(a) a polymer particle having an inner core and an outer shell wherein the inner core is a polymer having a refractive index of not less than 1.54 as measured at the wavelength of the sodium D line and wherein the outer shell is a polymer of

(i) an ethylenically unsaturated monomer having a functional group selected from the group consisting of epoxy, carboxyl, and aldehyde, said functional group capable of reacting with a hydrophilic polymeric linker; and

(ii) not more than 10 parts by weight of the outer shell of the monomers of the inner core, said outer shell being formed by polymerization in the presence of said inner core; and

wherein said polymer particle has an approximate diameter range of 0.03-0.1 μ

m;

covalently attached to

(b) the compound of biological interest through a hydrophilic polymeric linker of empirical formula

space="preserve" listing-type="equation">H[NHC.sub.2 H.sub.3 R(OC.sub.2 H.sub.3 R).sub.x OC.sub.2 H.sub.3 R].sub.y NH.sub.2

wherein R is at least one member selected from the group consisting of H and CH₃, x is 0-70 and y is 1-20,

(2) a liquid suspected of containing the compound of biological interest; and

(3) an agglutinating agent; and

(B) photometrically measuring increased particle size resulting from agglutination.

10. The method of claim 8 or claim 9 wherein R═

H.

11. The method of claim 10 wherein X=1-15 and y=1-15.

9. The particle reagent of claim 9 wherein said outer shell additionally contains other ethylenically unsaturated monomers in an amount sufficient to render the polymer particles substantially water insoluble.