IgG antibody testing method
First Claim
1. A method of determining if the concentration of IgG antibodies in biological fluids of a mammal is below a predetermined threshold level, the method comprising the steps of:
- taking a sample of a biological fluid from the mammal;
exposing a quantity of the sample to a quantity of a dehydrated IgG complexing agent coupled with a labeling agent that is immobilized at a sample position on a protein binding membrane to yield a conjugate;
exposing the conjugate to a quantity of a dehydrated standard mammalian IgG immobilized at a test position on the membrane, the test position being spaced from the sample position on the membrane; and
observing binding of the IgG complexing agent with the standard mammalian IgG evidenced by a color change from the labeling agent at a test position which color change indicates that the concentration of IgG antibodies is below the predetermined threshold level.
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Abstract
A method of determining the concentration of IgG antibodies in the biological fluids of mammals. The assay may be performed in a lateral flow cassette or dipstick format where dehydrated immobilized reagents are spaced along a membrane. A sample of a mammalian biological fluid is exposed first to an IgG complexing agent to yield a conjugate. The conjugate then moves along the membrane and is exposed to a standard mammalian IgG applied to the membrane at a test position. Binding of the IgG is indicated by a color change at the test position on the membrane. A control reagent of dehydrated anti-IgG complexing agent may be applied to the membrane at the control position spaced downstream from the test position. Interaction of the IgG complexing agent with the anti-IgG complexing agent forms a visible line at the control position on the membrane.
32 Citations
26 Claims
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1. A method of determining if the concentration of IgG antibodies in biological fluids of a mammal is below a predetermined threshold level, the method comprising the steps of:
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taking a sample of a biological fluid from the mammal;
exposing a quantity of the sample to a quantity of a dehydrated IgG complexing agent coupled with a labeling agent that is immobilized at a sample position on a protein binding membrane to yield a conjugate;
exposing the conjugate to a quantity of a dehydrated standard mammalian IgG immobilized at a test position on the membrane, the test position being spaced from the sample position on the membrane; and
observing binding of the IgG complexing agent with the standard mammalian IgG evidenced by a color change from the labeling agent at a test position which color change indicates that the concentration of IgG antibodies is below the predetermined threshold level. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
exposing the conjugate to a quantity of dehydrated anti-IgG complexing agent immobilized at a control position on the membrane, the control position being spaced from the test position; and
observing binding of the IgG complexing agent and the anti-IgG complexing agent evidenced by a color change from the labeling agent at a control position.
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3. The method of claim 1, further including the step of:
- diluting the sample before exposing it to the IgG complexing agent.
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4. The method of claim 2, further including the step of:
- diluting the sample before exposing it to the IgG complexing agent.
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5. The method of claim 1, wherein the IgG complexing agent and standard mammalian IgG are applied and dried in spaced relationship on a protein binding membrane.
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6. The method of claim 2, wherein the anti-IgG complexing agent is immobilized on a protein binding membrane.
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7. The method of claim 5, wherein the membrane is formed of a material selected from the group consisting of polyvinylidene difluoride, mixed cellulose ester, and nitrocellulose.
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8. The method of claim 6, wherein the membrane is formed of a material selected from the group consisting of polyvinylidene difluoride, mixed cellulose ester, and nitrocellulose.
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9. The method of claim 1, wherein the biological fluid is selected from a group consisting of whole blood, plasma, serum, colostrum, and urine.
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10. The method of claim 1, wherein the mammal is selected from a group consisting of cats, cattle, dogs, horses, humans and swine.
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11. The method of claim 1, wherein the predetermined threshold level ranges from about 5 mg/ml to about 15 mg/ml.
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12. The method of claim 11, wherein the predetermined threshold level is about 10 mg/ml.
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13. The method of claim 1, wherein the IgG complexing agent is selected from a group consisting of native Protein A, native Protein G, recombinant Protein A, recombinant Protein G, a monoclonal anti-IgG antibody, a polyclonal anti-IgG antibody, and mixtures thereof.
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14. The method of claim 1, wherein the IgG complexing agent is coupled with a labeling agent selected from a group consisting of colloidal gold and latex particles.
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15. The method of claim 5, wherein the membrane is supported in a lateral flow cassette, and wherein the sample is deposited on the membrane through an opening in the cassette.
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16. The method of claim 5, wherein the membrane is supported on a dipstick structure, and wherein the sample is deposited on the membrane by placing the dipstick structure into the sample.
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17. The method of claim 5, wherein a first quantity of standard mammalian IgG is immobilized at a first test position on the membrane, and a second quantity of standard mammalian IgG is immobilized at a second test position spaced from the first test position, wherein binding of the IgG complexing agent with the first and second quantities of standard mammalian IgG is evidenced by color changes at the first and second test positions.
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18. The method of claim 17, wherein a third quantity of standard mammalian IgG is immobilized at a third test position on the membrane, and wherein binding of the IgG complexing agent with the third quantity of standard mammalian IgG is evidenced by a color change at the third test position.
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19. A method of determining if the concentration of IgG antibodies in biological fluids of a mammal is below a predetermined threshold level, the method comprising the steps of:
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taking a sample of the biological fluid from the mammal;
exposing a quantity of the sample to a quantity of dehydrated IgG complexing agent coupled with a labeling agent that is immobilized at a sample position on a protein binding membrane to yield a conjugate;
exposing the conjugate to a quantity of dehydrated standard mammalian IgG immobilized at a test position on the membrane, the test position being spaced from the sample position on the membrane; and
observing binding of the IgG complexing agent with the standard mammalian IgG evidenced by a color change from the labeling agent at the test position, which color change indicates that the concentration of IgG antibodies is below the predetermined threshold level. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26)
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Specification