Cancer diagnosis using Ki-67
The present invention provides methods and compositions for detection, diagnosis, prognosis, monitoring treatment and monitoring progression of cancer by detecting the level of Ki-67 protein in an individual.
1 . A method comprising:
(a) obtaining a plasma sample from a subject suspected of having lymphoma;
(b) contacting the plasma sample with an agent that binds to a Ki-67 protein that is detectable in the plasma sample, wherein the plasma sample contains less than about 1% (w/w) whole cellular material; and
(c) detecting a level of Ki-67 protein in the plasma sample by performing an immunoassay, wherein the level of Ki-67 protein is above about 1500 U/mL.
2 . The method of claim 1 , wherein (a) comprises centrifuging the plasma sample.
3 . The method of claim 1 , wherein the agent is immobilized on a support.
4 . The method of claim 3 , wherein the support is a microwell, bead, or microarray.
5 . The method of claim 3 , wherein the support is nitrocellulose, glass, polyacrylamides, gabbros, or magnetite.
6 . The method of claim 3 , wherein the support is spherical, cylindrical, or flat.
7 . The method of claim 1 , wherein the agent is an anti-Ki-67 antibody.
8 . The method of claim 7 , wherein the anti-Ki-67 antibody is labeled with a detectable label.
9 . The method of claim 8 , wherein the detectable label is selected from the group consisting of a radioisotope, enzyme, enzyme substrate, luminescent substance, biotinyl group, and predetermined polypeptide epitope recognized by a secondary reporter.
10 . The method of claim 1 , wherein the Ki-67 protein comprises the amino acid sequence of SEQ ID NO: 1.
11 . The method of claim 1 , wherein the immunoassay is selected from a radioimmunoassay, immunoprecipitation, Western blot, enzyme-linked immunosorbent assay (ELISA), electrochemiluminescence assay, or 2-site or sandwich immunoassay.
12 . The method of claim 1 , wherein the lymphoma is non-Hodgkin's lymphoma (NHL) or Burkitt's lymphoma.
13 . The method of claim 1 , further comprising measuring a level of β2-microglobulin in the plasma sample.