Methods and compounds for lymphoma cell detection and isolation

Abstract

Compositions comprising a purified, isolated antibody, humanized antibodies and precipitates directed against ROR-1, wherein the antibody binds ROR-1 with moderate to high affinity. The compositions may be used for detecting and isolating an amount of ROR-1 in a subject sample, and to evaluate the appearance, status, course, or treatment of a ROR-1 cancer in a subject. The ROR-1 antibodies are especially useful in identifying lymphomas and ademocarcinomas. Vaccines and related methods for protecting a subject against diseases that involve expression of ROR-1 are also provided.

1 . A composition comprising a purified, isolated antibody directed against ROR-1, wherein the antibody binds ROR-1 with moderate to high affinity.  
   
   
       2 . A composition according to  claim 1 , wherein the antibody is an anti-ROR-1 polyclonal antibody, a monoclonal antibody, or a functional antibody fragment.  
   
   
       3 . A composition according to  claim 2 , wherein the antibody comprises a sequence expressed by a heavy chain sequence and at least one light chain sequence of SEQ ID NOs: 1-5.  
   
   
       4 . A composition according to  claim 2 , wherein the antibody is a polyclonal antibody.  
   
   
       5 . A composition according to  claim 2 , wherein the antibody is a monoclonal antibody.  
   
   
       6 . A composition according to  claim 2 , wherein the antibody is a functional antibody fragment.  
   
   
       7 . A composition according to  claim 1 , wherein the antibody is selected from the group consisting of whole antibody, humanized antibody, chimeric antibody, Fab fragment, Fab′ fragment, F(ab′) 2  fragment, single chain Fv fragment and diabody.  
   
   
       8 . A composition according to  claim 1 , wherein the antibody has an affinity to binding ROR-1 with a dissociation constant of below a Kd value selected from the group consisting of 10 −6  mol/l, 10 −7  mol/l, and 10 −8  mol/l.  
   
   
       9 . A composition according to  claim 1 , wherein the antibody is a detectably labeled antibody.  
   
   
       10 . A composition according to  claim 1 , further comprising a pharmaceutically acceptable agent.  
   
   
       11 . A method for detecting an amount of ROR-1 in a subject sample, the method comprising: 
 (a) contacting the subject sample with the anti-ROR-1 antibody of  claim 1;  and    (b) detecting immunoreactivity between the anti-ROR-1 antibody and ROR-1 in the sample.    
   
   
       12 . A method according to  claim 11 , wherein the antibody is covalently attached to a detectable label.  
   
   
       13 . A method according to  claim 11 , wherein the immunoreactivity detection is provided by immunoperoxidase staining, immunofluorescence, immunoelectronmicroscopy, or ELISA.  
   
   
       14 . A method according to  claim 11 , further comprising 
 (c) correlating binding of the antibody to a standardized antibody binding profile, wherein such correlation provides quantitative value for total CPR in the sample.    
   
   
       15 . A method for detecting a ROR-1 cancer, the method comprising: detecting the presence or quantity of ROR-1 protein in a subject sample.  
   
   
       16 . A method according to  claim 15 , wherein the ROR-1 cancer is a lymphoma or adeno carcinoma.  
   
   
       17 . A method according to  claim 16 , wherein the cancer is selected from the group consisting of CLL, small lymphocytic lymphoma, marginal cell B-Cell lymphoma, Burkett's Lymphoma, colon adenocarcinoma, colorectal adenocarcinoma, and breast adenocarcinoma.  
   
   
       18 . A method according to  claim 15 , wherein detection of ROR-1 comprises the method of  claim 10 .  
   
   
       19 . A method for treating a ROR-1 cancer in a subject, the method comprising: 
 administering to the subject in need thereof a therapeutically effective amount of a ROR-1 receptor agonist.    
   
   
       20 . A method according to  claim 19 , wherein the ROR-1 cancer is a lymphoma or adenocarcinoma.  
   
   
       21 . A method according to  claim 19 , wherein the lymphoma is selected from the group consisting of CLL, small lymphocytic lymphoma, marginal cell B-Cell lymphoma, and Burkett's Lymphoma, colon adenocarcinoma, and breast adenocarcinoma.  
   
   
       22 . A method according to  claim 19 , wherein the ROR-1 receptor agonist is the anti-ROR-1 antibody of  claim 1 .  
   
   
       23 . A method according to  claim 22 , wherein the antibody is administered in an amount of (i) about 0.05 mg to about 2.5 mg; (ii) about 0.1 mg to about 1 mg; or (iii) about 0.3 mg to about 0.5 mg.  
   
   
       24 . A method according to  claim 22 , wherein the anti-ROR-1 antibody is a polyclonal antibody, a monoclonal antibody, or a functional antibody fragment.  
   
   
       25 . A method according to  claim 22 , wherein the anti-ROR-1 antibody is selected from the group consisting of whole antibody, humanized antibody, chimeric antibody, Fab fragment, Fab′ fragment, F(ab′) 2  fragment, single chain Fv fragment and diabody.  
   
   
       26 . A method according to  claim 19 , wherein the ROR-1 receptor agonist is an antisense inhibitor of ROR-1.  
   
   
       27 . A method according to  claim 26 , wherein the ROR-1 antisense inhibitor is administered in an amount of (i) about 10 μg/day to about 3 mg/day; (ii) about 30 μg/day to about 300 μg/day; or (iii) about 100 μg/day.  
   
   
       28 . A method according to  claim 19 , wherein the ROR-1 receptor agonist is administered by injection, inhalation, orally, liposome, or retroviral vector.  
   
   
       29 . A diagnostic method for evaluating the appearance, status, course, or treatment of a ROR-1 cancer in a subject, the method comprising: 
 (a) contacting a subject sample with the anti-ROR-1 antibody of  claim 1;  and    (b) detecting immunoreactivity between the anti-ROR-1 antibody and ROR-1 to determine presence or quantity of ROR-1 in the sample.    
   
   
       30 . A method according to  claim 29 , wherein the antibody specifically binds to ROR-1.  
   
   
       31 . A method according to  claim 29 , wherein a diagnostic criterion or value is determined based on an increase or decrease in an amount of ROR-1 in the subject compared to a control level(s) of ROR-1 in a normal subject or sample.  
   
   
       32 . A method according to  claim 29 , wherein immunoreactivity detection is provided by immunoperoxidase staining, immunofluorescence, immunoelectronmicroscopy, or ELISA.  
   
   
       33 . A method according to  claim 29 , wherein the antibody is an anti-ROR-1 polyclonal antibody, a monoclonal antibody, or a functional antibody fragment.  
   
   
       34 . A method according to  claim 33 , wherein the antibody is selected from the group consisting of whole antibody, humanized antibody, chimeric antibody, Fab fragment, Fab′ fragment, F(ab′) 2  fragment, single chain Fv fragment and diabody.  
   
   
       35 . A kit to detect the presence of ROR-1 protein comprising the antibody of  claim 1 .  
   
   
       36 . A vaccine composition comprising a polynucleotide encoding ROR-1 protein or a fragment or variant thereof, and a pharmaceutically acceptable carrier or diluent.  
   
   
       37 . A vaccine composition comprising ROR-1 protein or a fragment or variant thereof, and a pharmaceutically acceptable carrier or diluent.  
   
   
       38 . A method for protecting against the occurrence of diseases involving expression of ROR-1 in a subject, the method comprising: 
 administering to the subject in need thereof a polynucleotide encoding ROR-1 protein or a fragment or variant thereof in an amount effective to induce a protective or therapeutic immune response against ROR-1 in the subject, and a pharmaceutically acceptable carrier or diluent.    
   
   
       39 . A method for protecting against the occurrence of diseases involving expression of ROR-1 in a subject, the method comprising: 
 administering to the subject in need thereof a ROR-1 protein or a fragment or variant thereof in an amount effective to induce a protective or therapeutic immune response against ROR-1 in the subject, and a pharmaceutically acceptable carrier or diluent.    
   
   
       40 . A method for identifying or isolating ROR-1 protein in a sample, the method comprising: 
 (a) contacting the sample with the anti-ROR-1 antibody of  claim 1;  and    (b) detecting immunoreactivity between the anti-ROR-1 antibody and ROR-1 to determine presence or quantity of ROR-1 in the sample.    
   
   
       41 . A method according to  claim 40 , wherein the ROR-1 antibody is conjugated to a magnetic bead.  
   
   
       42 . A method for detecting minimal residual disease following treatment of a ROR-1 cancer, the method comprising: 
 (a) contacting the sample with the anti-ROR-1 antibody of  claim 1;  and    (b) detecting immunoreactivity between the anti-ROR-1 antibody and ROR-1 to determine presence or quantity of ROR-1 in the sample.    
   
   
       43 . A method according to  claim 42 , wherein the ROR-1 antibody is conjugated to a magnetic bead.  
   
   
       44 . A humanized ROR-1 antibody.  
   
   
       45 . A precipitate comprising a ROR-1 antibody and a polypeptide selected from the group consisting of a ROR-1 protein, ROR-1 polypeptide fragment and ROR-1 polypeptide variant.