Multi epitope assay

1. A method for detecting prostate-specific antigen in an affinity assay, the method comprising the steps of:

a) providing a biological sample volume containing the prostate-specific antigen at a concentration of from at 4 to 10 ng/ml;

b) adding PSA-10 antibody capturing moiety to the biological sample volume, wherein the PSA-10 antibody capturing moiety captures the prostate-specific antigen and is adhered to a magnetic particle, wherein the PSA-10 antibody capturing moiety is linked to the particle via a linker comprising a DNA linker consisting of 48 base pairs of double stranded DNA followed by 10 bases of single stranded DNA, wherein the DNA is digestable by a DNAse and cleavable for specific elution with EcoRI;

c) concentrating the captured prostate-specific antigen by magnetic particle separations into an elution volume of less than 5 μL that is at least eighty times smaller than the biological sample volume in step a) in a time of fifteen minutes;

d) cleaving the prostate-specific antigen from the particle, wherein at least 30% of the prostate-specific antigen is cleaved from the particle using a DNAse or EcoRI;

e) binding the prostate-specific antigen to a modified PSA-36 antibody capturing moiety associated with a reagent and to a modified PSA-66 antibody capturing moiety comprising a tag, wherein a portion of the PSA-10 antibody capturing moiety that was cleaved from the particle binds to one of the modified antibodies, and

f) at least one of detecting and quantifying the prostate-specific antigen in an affinity assay format, wherein the tag binds to a fourth capturing moiety on a detection surface.

2. The method according to claim 1 , wherein the reagent is selected from the group consisting of;

a radioisotope,

a solid phase,

a fluorescent, phosphorescent, or chemiluminescent dye,

an enzyme or enzyme substrate,

a nucleic acid sequence or peptide,

a colloid or nanoparticle, and

a polymer or macromolecule.

3. The method according to claim 1 , wherein the at least one of detecting and quantifying step e) is performed according to a method selected from the group consisting of:

an optical detection,

an enzyme reaction,

nucleic-acid amplification techniques,

a magnetic detection,

a sonic detection, and

an electrical detection.

4. The method according to claim 1 , wherein the tag comprises biotin and the fourth capturing moiety comprises either avidin or streptavidin.

5. A method for detecting prostate-specific antigen in an affinity assay, the method comprising the steps of:

a) providing a biological sample volume containing the prostate-specific antigen at a concentration of from at 4 to 10 ng/ml;

b) adding PSA-10 antibody capturing moiety to the biological sample volume, wherein the PSA-10 antibody capturing moiety captures the prostate-specific antigen and is adhered to a magnetic particle, wherein the PSA-10 antibody capturing moiety is linked to the particle via a linker comprising a DNA linker consisting of 48 base pairs of double stranded DNA followed by 10 bases of single stranded DNA, wherein the DNA is digestable by a DNAse and cleavable for specific elution with EcoRI;

c) concentrating the captured prostate-specific antigen by magnetic particle separations into an elution volume of less than 5 μL that is at least eighty times smaller than the biological sample volume in step a) in a time of fifteen minutes;

d) cleaving the prostate-specific antigen from the particle, wherein at least 30% of the prostate-specific antigen is cleaved from the particle and wherein the cleavage is enzymatic;

e) binding the prostate-specific antigen to a modified PSA-36 antibody capturing moiety associated with a reagent and to a modified PSA-66 antibody capturing moiety comprising a tag, wherein a portion of the PSA-10 antibody capturing moiety that was cleaved from the particle binds to one of the modified antibodies, and

f) at least one of detecting and quantifying the prostate-specific antigen in an affinity assay format, wherein the tag binds to a fourth capturing moiety on a detection surface.

6. A method for detecting prostate-specific antigen in an affinity assay, the method comprising the steps of:

a) providing a biological sample volume containing the prostate-specific antigen at a concentration of from at 4 to 10 ng/ml;

b) adding PSA-10 antibody capturing moiety to the biological sample volume, wherein the PSA-10 antibody capturing moiety captures the prostate-specific antigen and is adhered to a magnetic particle, wherein the PSA-10 antibody capturing moiety is linked to the particle via a first entity for attachment to a particle comprising a DNA linker, wherein the DNA is digestable by a DNAse and cleavable for specific elution with EcoRI;

c) concentrating the captured prostate-specific antigen by magnetic particle separations into an elution volume of less than 5 μL that is at least eighty times smaller than the biological sample volume in step a) in a time of fifteen minutes;

d) cleaving the prostate-specific antigen from the particle, wherein at least 30% of the prostate-specific antigen is cleaved from the particle using a second entity for the cleavage of the attachment to the particle;

e) binding the prostate-specific antigen to a modified PSA-36 antibody capturing moiety associated with a reagent and to a modified PSA-66 antibody capturing moiety comprising a third entity for attachment to a detection surface, wherein a portion of the PSA-10 antibody capturing moiety that was cleaved from the particle binds to one of the modified antibodies, and

f) at least one of detecting and quantifying the prostate-specific antigen in an affinity assay format, wherein the third entity binds to a fourth capturing moiety on a detection surface.