Human antibodies as detection reagents
View Patent ↗The invention provides in vitro methods of detection using human antibodies. The methods are particularly useful for analyzing human samples containing HAMA or heterophilic antibodies. A human antibody can bind to an analyte in such samples without binding to HAMA or heterophilic antibodies present in the samples. The methods can be effected using a sandwich format among others.
1. A method of detecting an analyte, comprising:
contacting a sample with at least 100 human antibodies immobilized to one or more supports, each antibody having an affinity of at least 10 8 M −1 for a different analyte that may be present in the sample,
detecting binding of at least one of the human antibodies to the analyte for which it has the affinity indicating presence and/or amount of the analyte in the sample.
2. The method of claim 1 , wherein the affinity of the human antibodies for the analyte is at least 10 10 M −1 .
3. The method of claim 1 , wherein the affinity of the human antibody for the analyte is at least 10 11 M −1 .
4. The method of claim 1 , wherein the different human antibodies occupy different known locations on the same support.
5. The method of claim 4 , wherein the detecting comprises detecting binding of at least two of the antibodies to at least two analytes in the sample.
6. The method of claim 4 , wherein less than 500 ng of each human antibody is present in the known locations.
7. The method of claim 4 , wherein less than 100 ng of each human antibody is present in the known locations.
8. The method of claim 1 , wherein the different human antibodies are immobilized to a plurality of supports and the detecting step comprises detecting binding of at least two of the human antibodies to at least two analytes to indicate presence and/or amount of the analytes in the sample.
9. The method of claim 8 , wherein the plurality of human antibodies comprises at least 1000 different antibodies having specific affinities for different analytes that may be present in the sample.
10. The method of claim 1 , wherein the sample comprises an unlabelled form of one of the analytes to be detected and a labelled form of the analyte, and the detecting step detects binding of the human antibody to the labelled form of the analyte, the extent of binding being inversely related to the amount of the unlabelled analyte in the sample.
11. The method of claim 1 , wherein the sample is further contacted with a human antibody in solution, one of the immobilized human antibodies and the human antibody in solution binding to different epitopes of the same analyte, and the detecting step comprising detecting binding of the immobilized human antibody and the human antibody in solution to the analyte.
12. The method of claim 1 , wherein the sample is a mixture of a first sample bearing different analytes labelled with a first label and a second sample bearing different analytes labelled with a second label, the detecting comprises detecting a ratio of first label to second label bound to a plurality of antibodies, to indicate ratios of amounts of analytes present in the first and second samples.
13. The method of claim 1 , wherein the specific affinities of the different antibodies for the different analytes fall within a factor of ten of each other.
14. The method of claim 1 , wherein the sample is a sample from a human subject.
15. The method of claim 14 , wherein the sample contains HAMA and/or heterophilic antibodies.
16. The method of claim 1 , wherein the analyte is a human molecule.
17. A plurality of human antibodies each having an affinity of at least 10 8 M −1 for a different corresponding analyte, the different human antibodies occupying different known locations on one or more supports, wherein the plurality of antibodies is at least 100.
18. The plurality of human antibodies of claim 17 , wherein the different antibodies occupy different known locations of the same support.
19. The plurality of human antibodies of claim 18 , wherein the known locations have areas less than 0.01 cm 2 .
20. The plurality of human antibodies of claim 18 , wherein the known locations have areas less than 0.001 cm 2 .
21. The plurality of human antibodies of claim 18 , wherein the known locations have areas less than 0.0001 cm 2 .
22. The plurality of human antibodies of claim 18 , wherein the known locations have areas less than 0.00001 cm 2 .
23. The plurality of human antibodies of claim 18 , wherein the known locations have areas less than 0.0000001 cm 2 .
24. The plurality of antibodies of claim 17 , wherein each analyte is a human molecule.
25. The plurality of antibodies of claim 17 , wherein each of the antibodies has an affinity of at least 10 10 M −1 .
26. The plurality of antibodies of claim 17 , wherein each of the antibodies has an affinity of at least 10 11 M −1 .