ELISA detection of urine DEK to predict and diagnose bladder cancer in humans
The present invention is directed to a method of detecting a DEK protein in a human urine sample using an ELISA assay. Methods and compositions for detection of DEK using mAb 260-6F9F6 (as detection antibody) and mAb 16-2C9C3 (as capture antibody) in human urine are provided herein. Specifically, the ELISA assay utilizes a capture mAb and a detection mAb to yield a high sensitivity of <50 ng/mL. The presence of DEK in urine is useful in predicting or diagnosing the occurrence of bladder cancer in humans.
1. A method of detecting DEK protein in a biological sample, comprising the steps of:
a. providing a biological sample;
b. immobilizing an isolated monoclonal antibody (mAb 16-2C9C3) onto a solid surface,
c. said mAb 16-2C9C3 comprises: (i) CDR1 of the heavy chain variable region which has the amino acid sequence of SEQ ID NO:15; (ii) CDR2 of the heavy chain variable region which has the amino acid sequence of SEQ ID NO:16; (iii) CDR3 of the heavy chain variable region which has the amino acid sequence of SEQ ID NO:17; (iv) CDR1 of the light chain variable region which has an amino acid sequence of SEQ ID NO: 23; (v) CDR2 of the light chain variable region which has the amino acid sequence of SEQ ID NO:24; and (vi) CDR3 of the light chain variable region which has the amino acid sequence of SEQ ID NO:25;
d. adding said biological sample onto said solid surface to allow DEK protein present in said biological sample to be captured onto said solid surface by said isolated monoclonal antibody (mAb 16-2C9C3);
e. washing the solid surface to remove unbound DEK protein;
f. adding an isolated monoclonal antibody (mAb 260-6F9F6) so as to allow formation of a complex between said captured DEK protein with said isolated monoclonal antibody (mAb 260-6F9F6), said mAb 260-6F9F6 comprises: (i) CDR1 of the heavy chain variable region which has the amino acid sequence of SEQ ID NO:31; (ii) CDR2 of the heavy chain variable region which has the amino acid sequence of SEQ ID NO:32; (iii) CDR3 of the heavy chain variable region which has the amino acid sequence of SEQ ID NO:33; (iv) CDR1 of the light chain variable region which has an amino acid sequence of SEQ ID NO: 39; (v) CDR2 of the light chain variable region which has the amino acid sequence of SEQ ID NO:40; and (vi) CDR3 of the light chain variable region which has the amino acid sequence of SEQ ID NO:41; and
g. detecting said complex,
wherein the presence of said complex is indicative of the presence of DEK in said biological sample; and
wherein said biological sample is urine.
2. A method of detecting DEK protein in a biological sample, comprising the steps of:
a. providing a biological sample;
b. immobilizing an isolated monoclonal antibody (mAb 16-2C9C3) onto a solid surface,
said mAb 16-2C9C3 comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 14 and a light chain variable region having the amino acid sequence of SEQ ID NO: 22;
c. adding said biological sample onto said solid surface to allow DEK protein present in said biological sample to be captured onto said solid surface by said isolated monoclonal antibody (mAb 16-2C9C3);
d. washing the solid surface to remove unbound DEK protein;
e. adding an isolated monoclonal antibody (mAb 260-6F9F6) so as to allow formation of a complex between said captured DEK protein with said isolated monoclonal antibody (mAb 260-6F9F6),
said mAb 260-6F9F6 comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 30 and a light chain variable region having the amino acid sequence of SEQ ID NO: 38; and
f. detecting said complex,
wherein the presence of said complex is indicative of the presence of DEK in said biological sample; and
wherein said biological sample is urine.