Novel forms of prostate specific antigen (PSA) specific for benign prostatic hyperplasia (BPH) and methods of using such

Abstract

A substantially pure and isolated novel form of prostate specific antigen (PSA) is provided. The novel form of PSA of the present invention comprises at least one clip at Lys 182 of the amino acid sequence of a mature form of PSA. Preferably, the novel form of PSA additionally comprises one or more clips at a location selected from a group consisting of Ile 1, Lys 145, and Lys 146. More preferably, the form of PSA contains at least two clips at Lys 145 and Lys 182 of the amino acid sequence of a mature form of PSA. The novel forms of PSA exist at an elevated level in patients suspected of having benign prostatic hyperplasia (BPH) and therefore may be used as a serum mark or an immunohistological marker to help distinguish BPH from prostate cancer. Antibodies recognizing the novel forms of PSA and immunoassays that detect and determine the novel forms of PSA of the present invention in a sample are also provided. Further provided are a kit and a method for detecting the novel forms of PSA for aiding in the differentiation of prostate cancer from BPH.

What is claimed is:  
     
         1 . A substantially pure and isolated form of prostate specific antigen (PSA) comprising at least one clip at lysine 182 of the amino acid sequence of a mature form of PSA.  
     
     
         2 . The substantially pure and isolated form of PSA of  claim 1  additionally comprising one or more clips at a location selected from a group consisting of Ile 1, Lys 145, and Lys 146.  
     
     
         3 . The substantially pure and isolated form of PSA of  claim 2  consisting of two clips at Lys 145 and Lys 182 of the mature form of PSA.  
     
     
         4 . The PSA of  claim 1  existing at an elevated level in transition zone prostate tissue.  
     
     
         5 . The PSA of  claim 1  existing as a component of free PSA in BPH.  
     
     
         6 . The PSA of  claim 3  being separable from other forms of free PSA by HIC-HPLC technique.  
     
     
         7 . An antibody that is preferentially reactive with a form of PSA that comprises at least one clip at Lys 182 of the amino acid sequence of a mature form of PSA.  
     
     
         8 . The antibody of  claim 7 , wherein the form of PSA additionally comprises one or more clips at a location selected from a group consisting of Ile 1, Lys 145, and Lys 146.  
     
     
         9 . The antibody of  claim 8 , wherein the form of PSA consists of two clips at Lys 145 and Lys 182 of the amino acid sequence of a mature form of PSA.  
     
     
         10 . The antibody of  claim 7 , wherein the antibody is a polyclonal antibody.  
     
     
         11 . The antibody of  claim 7 , wherein the antibody is a monoclonal antibody.  
     
     
         12 . The antibody of  claim 11 , wherein the monoclonal antibody is selected from a group consisting of PS2C109, PS2C501, PS2C634, PS2C807, and PS2C837.  
     
     
         13 . An immunogen that is capable of eliciting a monoclonal antibody which preferentially binds to a novel form of PSA comprising at least one clip at Lys 182 of the amino acid sequence of a mature form of PSA.  
     
     
         14 . A hybridoma cell line capable of producing a monoclonal antibody that preferentially binds a novel form of PSA comprising at least one clip at Lys 182 of the amino acid sequence of a mature form of PSA.  
     
     
         15 . The hybridoma cell line of  claim 14 , wherein the monoclonal antibody is selected from a group consisting of monoclonal antibodies PS2C109, PS2C501, PS2C634, PS2C807, and PS2C837.  
     
     
         16 . A method of producing monoclonal antibodies which preferentially binds to a novel form of PSA comprising at least one clip at Lys 182 of a mature form of PSA, the method comprising steps of: 
 (a) immunizing a mouse with a novel form of PSA; and    (b) producing monoclonal antibodies from the immunized mouse by employing a monoclonal antibody technique, wherein the monoclonal antibodies preferentially bind to the novel form of PSA compared to other forms of free PSA.    
     
     
         17 . The method of  claim 16 , wherein the monoclonal antibody produced by step (b) is selected from a group consisting of PS2C109, PS2C501, PS2C634, PS2C807, and PS2C837.  
     
     
         18 . A method of detecting, or determining in a sample, a form of prostate specific antigen (PSA) comprising at least one clip at lysine 182 of the amino acid sequence of a mature form of PSA, the method comprising the steps of: 
 (a) contacting an amount of an agent which specifically binds to the form of PSA to be detected with the sample under a condition that allows the formation of a binary complex comprising the agent and the form of PSA; and    (b) detecting or determining the presence or amount of the complex.    
     
     
         19 . The method of  claim 18 , wherein the sample is a sample of human physiological fluid.  
     
     
         20 . The method of  claim 19 , wherein the human physiological fluid is serum, seminal plasma, urine or plasma.  
     
     
         21 . The method of  claim 18 , wherein the agent is an antibody.  
     
     
         22 . The method of  claim 21 , wherein the antibody is a monoclonal antibody.  
     
     
         23 . The method of  claim 22 , wherein the monoclonal antibody is selected from a group consisting of PS2C109, PS2C501, PS2C634, PS2C807, and PS2C837.  
     
     
         24 . The method of  claim 19 , wherein the agent is an antibody and wherein the antibody is attached to a solid phase.  
     
     
         25 . The method of  claim 19 , wherein the agent is an antibody and wherein in step (b) the antibody comprises a detectable label or binds to a detectable label to form a detectable ternary complex.  
     
     
         26 . The method of  claim 18 , wherein the sample is a mammalian tissue sample.  
     
     
         27 . The method of  claim 26 , wherein the agent is an antibody.  
     
     
         28 . The method of  claim 26 , wherein in step (b), the complex is detected by a second agent which comprises a detectable label or which binds to a detectable label to form a detectable ternary complex.  
     
     
         29 . The method of  claim 28 , wherein the second agent is an antibody.  
     
     
         30 . The method of  claim 26 , wherein the mammalian tissue is a human prostate tissue.  
     
     
         31 . The method of  claim 18 , wherein the form of PSA additionally comprises one or more clips at a location selected from a group consisting of Ile 1, Lys 145, and Lys 146.  
     
     
         32 . The method of  claim 18 , wherein the form of PSA consists of two clips at Lys 145 and Lys 182 of the amino acid sequence of a mature form of PSA.  
     
     
         33 . A test kit for detecting or determining in a sample the form of PSA comprising at least one clip at lysine 182 of the amino acid sequence of a mature form of PSA, the kit comprising a known amount of an agent which preferentially binds to the form of PSA to be detected, wherein the agent is detectably labeled or binds to a detectable label.  
     
     
         34 . The test kit of  claim 33 , wherein the sample is a sample of human physiological fluid.  
     
     
         35 . The test kit of  claim 33 , wherein the agent comprises an antibody that preferentially binds to the form of PSA.  
     
     
         36 . The test kit of  claim 35  further comprising a solid phase capable of having the antibody attached thereto.  
     
     
         37 . The test kit of  claim 33 , wherein the sample is a mammalian tissue sample.  
     
     
         38 . The test kit of  claim 37 , wherein the agent is an antibody.  
     
     
         39 . The test kit of  claim 38 , wherein the antibody is a monoclonal antibody.  
     
     
         40 . The test kit of  claim 33 , wherein the agent is a monoclonal antibody selected from a group consisting of PS2C109, PS2C501, PS2C634, PS2C807, and PS2C837.  
     
     
         41 . The test kit of  claim 33 , wherein the form of PSA additionally comprises one or more clips at a location selected from a group consisting of Ile 1, Lys 145, and Lys 146.  
     
     
         42 . The test kit of  claim 41 , wherein the form of PSA consists of two clips at Lys 145 and Lys 182 of the amino acid sequence of a mature form of PSA.  
     
     
         43 . A method for distinguishing benign prostate hyperplasia from prostate cancer comprising: 
 (a) contacting an amount of an agent, which preferentially binds to a form of PSA comprising at least one clip at lysine 182 of the amino acid sequence of a mature form of PSA, with a sample obtained from a human containing the form of PSA under a condition sufficient to allow the formation of a binary complex comprising the agent and the form of PSA; and    (b) determining the amount of the complex in the sample and correlating the amount of the complex to the presence or absence of benign prostatic hyperplasia or prostate cancer in the human.    
     
     
         44 . The diagnostic method of  claim 43 , wherein the form of PSA additionally comprises one or more clips at a location selected from a group consisting of Ile 1, Lys 145, and Lys 146.  
     
     
         45 . The method of  claim 50 , wherein the form of PSA comprises two clips at Lys 145 and Lys 182 of the amino acid sequence of a mature form of PSA.  
     
     
         46 . The method of  claim 43 , wherein the sample is a sample of physiological fluid.  
     
     
         47 . The method of  claim 43 , wherein the sample is a mammalian tissue sample.  
     
     
         48 . The method of  claim 43 , wherein the agent is an antibody.  
     
     
         49 . The diagnostic method of  claim 48 , wherein the agent is a monoclonal antibody.  
     
     
         50 . The method of  claim 49 , wherein the monoclonal antibody is selected from a group consisting of PS2C109, PS2C501, PS2C634, PS2C807, and PS2C837.