Methods and apparatus for protein assay diagnostics
Automated protein assay apparatus and methods for measuring antibodies against an analyte are described. A standard mixture of one or more analytes is loaded into a capillary, the analytes are resolved by isoelectric focusing and immobilized in the capillary. Serum from a human or non-human subject under analysis is flowed through the capillary and antibodies specific for the immobilized analytes bind to the analytes. A secondary antibody including a detectable marker is introduced, binding to the immobilized antibody-analyte complexes. The locations of the antibody-analyte complexes are detected by means of the detectable markers, revealing the presence of analyte-specific antibodies in the serum.
1 . A method for performing a protein assay comprising:
loading a biological sample of one or more known analytes into a capillary;
resolving analytes of the sample in the capillary;
immobilizing the resolved analytes in the capillary;
binding patient antibodies to the immobilized analytes; and
detecting the location of antibody-analyte complexes.
2 . The method of claim 1 , wherein the patient antibodies are contained in patient serum and wherein loading is preceded by performing a screening test on the patient serum.
3 . The method of claim 2 , wherein performing a screening test comprises performing an ELISA assay.
4 . The method of claim 2 , wherein detecting further comprises detecting the location of patient antibodies which are particular to HIV.
5 . The method of claim 4 , wherein detecting further comprises detecting the locations of a plurality of patient antibodies which are particular to a plurality of proteins associated with HIV.
6 . The method of claim 2 , wherein detecting further comprises detecting the location of patient antibodies which are particular to Lyme disease.
7 . The method of claim 1 , wherein resolving comprises resolving analytes in the capillary by isoelectric focusing.
8 . The method of claim 7 , wherein detecting is preceded by flowing a chemiluminescent substrate through the capillary.
9 . The method of claim 1 wherein the steps of resolving, immobilizing, binding and detecting are performed by an integrated automated computer-controlled assay system.
10 . The method of claim 9 , wherein the integrated automated computer-controlled assay system further performs the step of loading.
11 . An automated assay system for detecting the presence of antibodies to a disease condition comprising:
a processing station at which one or more known resolved proteins are immobilized in a capillary;
a capillary gripper which is operable to manipulate capillaries at the processing station; and
a detection station, wherein the capillary gripper is further operable to manipulate processed capillaries containing a patient sample at the detection station,
wherein the presence of patient antibodies to one or more of the known proteins is detected at the detection station.
12 . The automated assay system of claim 11 , wherein the capillary gripper further comprises an automated, computer-controlled capillary gripper.
13 . The automated assay system of claim 12 , wherein the capillary gripper is operable to mechanically grip a capillary.
14 . The automated assay system of claim 12 , wherein the capillary gripper is operable to mechanically grip a capillary by a vacuum.
15 . The automated assay system of claim 11 , wherein the capillary gripper is further operable to load a biological sample into a capillary at the processing station.
16 . The automated assay system of claim 11 , further comprising a microtiter plate located at the processing station.
17 . The automated assay system of claim 11 , further comprising a photon detector located at the detection station.
18 . The automated assay system of claim 11 further comprising a user interface for configuring a protocol by which the automated assay system is to analyze the biological sample.
19 . The automated assay system of claim 18 , wherein the user interface is also operable to display detection results.
20 . A method for performing an assay comprising:
resolving pathogen-derived analytes in a capillary;
immobilizing the resolved analytes in the capillary;
loading patient-derived serum into the capillary;
allowing binding of patient-serum-derived antibodies to the immobilized analytes; and
detecting the location of serum-derived antibodies by means of a detection agent.
21 . The method of claim 20 , wherein resolving is preceded by performing a screening test on the patient-derived serum.
22 . The method of claim 21 , wherein performing a screening test comprises performing an ELISA assay.
23 . The method of claim 21 , wherein detecting further comprises detecting the location of analytes which are particular to HIV.
24 . The method of claim 21 , wherein detecting further comprises detecting the location of analytes which are particular to Lyme disease.
25 . The method of claim 20 , wherein resolving comprises resolving analytes in the capillary by isoelectric focusing.
26 . The method of claim 25 , wherein allowing binding comprises allowing antibodies to bind to the immobilized analytes.
27 . The method of claim 26 , wherein detecting is preceded by flowing a chemiluminescent substrate through the capillary.
28 . The method of claim 20 wherein the steps of resolving, immobilizing, loading and detecting are performed by an integrated automated computer-controlled assay system.
29 . The method of claim 20 , wherein resolving and immobilizing are performed prior to the time that a patient-derived serum is assayed,
whereby the patient-derived serum assay begins with a capillary pre-loaded with pre-resolved and pre-immobilized analytes.
30 . A method for performing an assay comprising:
resolving analytes in a capillary;
immobilizing the resolved analytes in the capillary;
loading serum into the capillary;
allowing serum-derived antibodies to bind to the immobilized analytes; and
detecting the location of serum-derived antibodies by means of a detection agent.
31 . The method of claim 30 , wherein the serum comprises human biological material.
32 . The method of claim 30 , wherein the serum comprises non-human biological material.
33 . The method of claim 30 , further comprising introducing a secondary antibody to which a detectable marker has been attached prior to detecting.
34 . The method of claim 33 , wherein allowing serum-derived antibodies to bind produces serum-derived antibody-analyte complexes; and
wherein the secondary antibody comprises an antibody which binds to the serum-derived antibody-analyte complexes.
35 . The method of claim 33 , further comprising washing prior to introducing a secondary antibody.
36 . The method of claim 30 , wherein resolving analytes further comprises resolving one or more known analytes in a capillary.
37 . The method of claim 36 , wherein resolving one or more known analytes further comprises resolving at least one of whole cells, viral particles, purified naturally occurring proteins, heterologously expressed proteins, or synthetic peptides.
38 . The method of claim 30 , wherein detecting further comprises detecting by one of chemiluminescence or fluorescence detection.
39 . The method of claim 30 , wherein resolving comprises resolving a plurality of analytes of a standard in a capillary;
wherein immobilizing comprises immobilizing the plurality of analytes at different locations in the capillary;
wherein loading serum comprises loading serum including a plurality of antibodies to the plurality of analytes into the capillary; and
wherein detecting comprises detecting multiple locations of serum-derived antibodies.
40 . The method of claim 30 wherein the steps of resolving, immobilizing, allowing serum-derived antibodies to bind, and detecting are performed by an integrated automated computer-controlled assay system.
41 . The method of claim 30 , further comprising:
recording results from the detection.
42 . The method of claim 41 wherein the steps of loading, allowing to bind, detecting and recording are performed by an integrated automated computer-controlled assay system.