Microarrays and uses therefor

• US 8,012,703 B2
• Filed: 06/23/2009
• Issued: 09/06/2011
• Est. Priority Date: 02/04/1998
• Status: Expired due to Fees

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Fernandez, Joseph, Hoeffler, James, Nasoff, Marc
• Primary Examiner(s): Cook; Lisa V
• Application Number:
  US12/490,069
• Publication Number:
  US 20100022407A1
• Time in Patent Office: 805 Days
• Field of Search: None
• US Class Current: 435/7.21
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/548 Carbohydrates, e.g. dextran
• CPC Class Codes: G01N 33/6845 Methods of identifying protein-protein interactions in protein mixtures
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

Methods of using microarrays to simplify analysis and characterization of genes and their function are provided. Such methods can be used to identify and characterize antibodies having binding affinity for a specific target antigen. A method of determining gene expression at the protein level by contacting an array of characterized or uncharacterized antibodies on a solid surface with one or more proteins and identifying the antibodies to which said protein(s) binds also is provided. This method can be used to compare the protein expression in two different populations of cells, such as normal cells and cancer cells or resting cells and stimulated cells. In addition, a method of determining gene expression at the protein level by contacting a microarray of nucleic acid samples derived from a variety of different sources with one or more nucleic acid probes then identifying the sample or samples to which the probe binds is provided.

Claims

1. A method of comparing protein expression in two or more populations of cells, said method comprising:

(a) contacting an array of antibodies bound to a solid surface with a cell lysate of a first cell population, generating a first binding pattern;

(b) contacting a duplicate array of antibodies on a solid surface with a cell lysate of a second cell population, generating a second binding pattern; and

(c) comparing the binding pattern of the first cell lysate with the binding pattern of the second cell lysate.

2. The method according to claim 1, wherein the antibodies are of uncharacterized binding specificity.

3. The method according to claim 1, wherein the antibodies are recombinant antibodies.

4. The according to claim 1, wherein the first cell lysate is derived from normal cells and the second cell lysate is derived from abnormal cells.

5. The method according to claim 4, wherein the abnormal cells are cancer cells.

6. The method according to claim 1, wherein the first cell lysate is derived from cells in a resting state and the second cell lysate is derived from cells in a stimulated state.

7. The method according to claim 1, wherein at least one of the one of the two cell lysates comprises a detectable label.

8. The method according to claim 1, wherein each of the two cell lysates comprises a different detectable label.

9. A method of comparing protein expression in two or more protein samples, said method comprising:

(a) contacting an array of antibodies bound to a solid surface with a first protein sample, generating a first binding pattern;

(b) contacting a duplicate array of antibodies on a solid surface with a second protein sample, generating a second binding pattern; and

(c) comparing the binding pattern of the first protein sample with the binding pattern of the second protein sample.

10. The method according to claim 9, wherein the first and second protein samples are derived from a body fluid.

11. The method according to claim 9, wherein the first and second protein samples are derived from serum samples.

12. The method according to claim 9, wherein the first and second protein samples are derived from cerebral spinal fluid, blood, plasma, urine, feces, saliva, tears, or extracted tissue.

13. The method according to claim 9, wherein the first and second protein samples are derived from cellular extracts.