Polymer particles containing colloidal iron oxide granules for use as a magnetically responsive reagent carrier

• US 5,206,159 A
• Filed: 08/25/1992
• Issued: 04/27/1993
• Est. Priority Date: 11/01/1984
• Status: Expired due to Fees

• Current Assignee: Miles, Inc. (Bayer AG)
• Original Assignee: Miles, Inc. (Bayer AG)
• Inventors: Cohen, Beri, Hargitay, Bartholomew, Wong, Tak K.
• Primary Examiner(s): Naff, David M.
• Application Number:
  US07/934,287
• Time in Patent Office: 245 Days
• Field of Search: 435/174, 435/176, 435/180, 435/4, 435/7.92, 436/526, 436/531, 436/532, 436/533, 436/534, 252/62.54, 252/62.56
• US Class Current: 435/180
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: C12N 11/14 Enzymes or microbial cells immobilised on or in an inorganic carrier
• CPC Class Codes: G01N 2446/10 the magnetic material being used to coat a pre-existing polymer particle but not being present in the particle core
• CPC Class Codes: G01N 2446/20 the magnetic material being present in the particle core
• CPC Class Codes: G01N 33/5434 using magnetic particle immunoreagent carriers which constitute new materials per se
• CPC Class Codes: G01N 33/545 Synthetic resin

Abstract

Substantially spherical polymer particles containing a colloidally, stably and uniformly dispersed magnetically responsive substance of about 5 nm to about 500 nm in diameter provide a superparamagnetic reagent carrier. Preferably, the polymer particles have a hydrated diameter of about 0.5 to 100 um, the polymer is a non-ionic cross-linked polyacrylamide gel and the magnetically responsive substance is iron oxide granules. The magnetically responsive substance may be present in an amount so that the polymer particles have a specific gravity of between 1.2 and 2.7. The reagent carrier is prepared by swelling polymer particles in a solution of iron salt so that substantially all of the iron salt solution is taken upon by the polymer particles and then converting the iron salt in situ to insoluble colloidally dispersed iron oxide granules. Converting of the iron salt may be carried out by reacting the iron salts with a base to form iron hydroxide and subsequently converting the hydroxide by heating to iron oxide. The polymer particles are particularly useful as a reagent carrier in automated immunoassays. Enzymes, antibodies, antigens or haptens may e coupled to the polymer particles.

Claims

1. A process for preparing a stable superparamagnetic particulate reagent carrier having high magnetic permeability and low magnetic remanence comprising the steps of:

a) providing a water-insoluble swellable substantially spherical and essentially non-ionic polyacrylamide gel matrix;

b) contacting said polyacrylamide matrix with a solution of soluble iron salts whereby substantially all of said iron salt solution is taken up by said polyacrylamide matrix, said matrix becomes swollen, and said soluble salts pass into said polyacrylamide hel matrix;

c) converting the iron salts to insoluble iron oxide granules having a diameter of from about 5 nm to 500 nm whereby a colloidally and stably dispersed magnetically responsive moiety is generated in situ within said polyacrylamide gel matrix and entrapped therein, said entrapped iron oxide essentially free to rotate within said matrix under the influence of a magnetic field and thermal motion; and

d) washing the polyacrylamide gel matrix to remove soluble by-products.

2. The process of claim 1, wherein the step of converting said iron salts comprises reacting said iron salts with a base to form iron hydroxides and subsequently converting said iron hydroxides by heating to iron oxide granules.

3. The process of claim 1, comprising the additional steps of:

e) drying the polyacrylamide matrix containing said iron oxide granules; and

f) heating them to elevated temperatures to increase the cross-link density of the polyacrylamie matrix, thereby reducing their swelling capacity when exposed to water or aqueous solutions.

4. A stable superparamagnetic regent carrier prepared by the process of claim 1.

5. The reagent carrier of claim 4, wherein said iron oxide granules are dispersed in the matrix in sufficient quantity to impart a specific gravity of about between 1.2 and 2.7 to said reagent carrier.

6. The reagent carrier of claim 4, wherein the colloidally dispersed iron oxide granules have a diameter smaller tan about 50 nm.

7. The reagent carrier of claim 4, wherein said iron oxide granules are at lest about 20% by weight of said reagent carrier.

8. The reagent carrier of claim 4, wherein a reagent component is immobilized on the surface of said matrix.