Antibody cocktail
View Patent ↗The present invention relates to a composition comprising at least three primary antibodies or fragments thereof, wherein the at least three antibodies or fragments thereof binds specifically to at least three different proteins, and wherein the at least three different proteins are AMCAR, CK 5/6, and HMWC. Methods for using the composition in diagnosis, prognosis, and assessing efficacy of treatment is further included as well as kits comprising said composition, and optionally, instructions of its use.
1. A composition for detecting three different proteins, AMCAR, CK 5/6, and HMWCK, comprising three different primary antibodies or antigen-binding fragments thereof that bind specifically to the three different proteins, respectively, wherein the antibody recognizing AMCAR is an antibody produced by a rabbit clone 13H4 or an antigen-binding fragment thereof, wherein the antibody recognizing CK 5/6 is an antibody produced by a mouse clone D5/16 B4 or an antigen-binding fragment thereof, and wherein the antibody recognizing HMWCK is an antibody produced by a mouse clone 34βE12 or an antigen-binding fragment thereof, and wherein no additional primary antibodies or antigen-binding fragments thereof are present in the composition.
2. The composition according to claim 1 , wherein at least one of the primary antibodies is a recombinant antibody.
3. The composition according to claim 1 , further comprising a buffer.
4. A kit comprising:
a. the composition of claim 1 , and
b. optionally, instructions for using the composition.
5. A method for in vitro detecting three different proteins in a biological sample, wherein the three different proteins are AMCAR, CK 5/6, and HMWCK, the method comprising the steps of
a. contacting said sample with the composition of claim 1 for a sufficient time to form three different antigen-antibody complexes, and
b. detecting said three different antigen-antibody complexes.
6. The method according to claim 5 , further comprising a step of optionally scoring the amount of antigen-antibody complexes.
7. The method according to claim 5 , further comprising step d) comparing the expression of said three proteins to at least one of a positive control and a negative control.
8. The method according to claim 5 , which is performed on an automated staining device.
9. The method according to claim 5 , wherein the detection is made manually.
10. The method according to claim 5 , wherein the detection is made by image analysis.
11. The method according to claim 5 , wherein the positive control comprises cells from a subject who is suffering from the prostate cancer, prostatic intraepithelial neoplasia (PIN), or benign mimics of prostate cancer.
12. The method according to claim 5 , wherein the negative control comprises cells from a healthy subject who is not suffering from prostate cancer, prostatic intraepithelial neoplasia (PIN), or benign mimics of prostate cancer.
13. The method of claim 7 , wherein the positive control comprises the proteins AMCAR, CK 5/6, and HMWCK.
14. The method of claim 7 , wherein the negative control does not comprise the proteins AMCAR, CK 5/6, or HMWCK.
15. The method of claim 5 , wherein the sample is obtained from a human prostate tissue sample.
16. The method of claim 15 , wherein the prostate tissue sample is obtained from a subject that has, is suspected of having, or previously had prostate cancer, prostatic intraepithelial neoplasia (PIN), or benign mimics of prostate cancer.
17. A method for producing an antibody-labeled prostate sample, comprising:
(i) providing a biological sample obtained from a sample of prostate tissue obtained from subject, and
(ii) contacting the biological sample with the composition of claim 1 ;
wherein the antibodies or antigen-binding fragments thereof bind to the sample to produce an antibody-labeled prostate sample.
18. The method of claim 17 , further comprising detecting the bound antibodies or antigen-binding fragments thereof.