Devices and methods for obtaining mammary fluid samples for evaluating breast diseases, including cancer

Abstract

Biological samples of mammary fluid or components thereof are obtained using a breast pump device coupled with a solid phase sample collection medium, alternatively facilitated by administering oxytocin to the subject. The breast pump device stimulates expression of mammary fluid and provides for collection of diagnostic samples to evaluate breast disease, including cancer. The biological sample may include whole cells or cellular components, purified or bulk proteins, glycoproteins, peptides, nucleotides or other desired constituents comprising a breast disease marker. Methods, kits and adapter devices relating to the breast pump device are also provided. Yet additional methods, devices, accessories, and materials are provided for laboratory handling and processing of breast fluid samples and for related diagnostic methods.

1 - 54 . (canceled)  
   
   
       55 . A sample collection device for collecting a biological sample from a mammary organ of a subject, comprising: 
 a breast engaging member for establishing a fluid connection between the engaging member and a nipple or alveolar duct of the subject;    pressure changing means connected with said breast engaging member for changing pressure at a surface of the nipple or within the alveolar duct to induce or facilitate breast fluid expression from the nipple or alveolar duct; and    a solid phase sample collection medium in fluid connection with said breast engaging member interposed between said breast engaging member and said pressure changing means for receiving a sample of expressed breast fluid from the nipple or alveolar duct, wherein said sample comprises one or more breast disease marker(s) selected from the group consisting of a protein, a peptide, a glycoprotein, a lipid, a glycolipid, a DNA polynucleotide, an RNA polynucleotide, or a combination thereof.    
   
   
       56 . The sample collection device of  claim 55 , wherein said biological sample comprises one or more breast disease marker(s) selected from the group consisting of Her 2, Ki67 Growth Factor, Cyclin BI, Cyclin D1, Proliferating Cell Nuclear Antigen, Transforming Growth Factor a, Tissue Plasminogen Activator, Insulin Growth Factor Receptors, Collagenase Type IV, Laminins, Laminin Receptor, Integrins, p53, rb, nm23, ras, c-myc, c-myb, Heat Shock Proteins, Prolactin, Neuron-Specific Enolase, IR-14, KA 1, KA 14, Alpha-Lactalbumin, Actin, CEA, HMFG, MCA, PSA, Vasopressin, Cathepsin D, PGE2, pS2; IL-10, S-100 protein; Vimentin; Epithelial Membrane Antigen, bcl-2, CA15-3, CA 19-9, Tn Antigen, Alpha-lactalbumin, LASA, Gal-GalNAC, GCDFP-15, Le(y)-Related Carbohydrate Antigen, CA 125, uPA, uPA related antigens and complexes, uPA Receptor, PA1-1 and PA1-2, Beta-glucuronidase, CD31, CD44 splice variants, blood group antigens including ABH, Lewis, and MN, and genetic lesions or altered expression levels of CCND1, EMS1, BRCA1 and BRCA2 genes, and combinations thereof.  
   
   
       57 . The sample collection device of  claim 55 , wherein said solid phase sample collection medium is in fluid contact with said nipple or alveolar duct when the breast engaging member is applied to the breast and positive or negative pressure is generated by said pressure changing means.  
   
   
       58 . The sample collection device of  claim 55 , wherein said pressure changing means is provided by said solid phase sample collection medium which operates by wicking or capillary action in contact with the nipple or alveolar duct to facilitate expression or evacuation of the sample of expressed breast fluid there from.  
   
   
       59 . The sample collection device of  claim 55 , wherein said solid phase sample collection medium is removably placed in fluid connection with a breast pump.  
   
   
       60 . The sample collection device of  claim 55 , wherein said solid phase sample collection medium is selected from the group consisting of microscopic glass slides, capillary tubes, collection tubes, columns, micro-columns, wells, plates, membranes, filters, resins, inorganic matrices, beads, resins, particulate chromatographic media, plastic microparticles, latex particles, coated tubes, coated templates, coated beads, coated matrices, and combinations thereof.  
   
   
       61 . The sample collection device of  claim 55 , wherein said biological sample is selected from the group consisting of whole mammary fluid, whole cells, cell fragments, cell membranes, proteins, glycoproteins, peptides, nucleotide components of mammary fluid, or a combination thereof.  
   
   
       62 . The sample collection device of claim  102 , wherein said pressure changing means functions to induce or facilitate expulsion of breast fluid from the nipple or alveolar duct independently of, or in conjunction with, positive pressure generated within the alveolar duct of the breast by spontaneous or oxytocin-induced contraction, or manual compression, of the alveolar duct.  
   
   
       63 . A non-invasive method for obtaining a biological sample from a mammary organ of a subject, comprising the steps of: 
 applying a sample collection device for collecting said biological sample from said subject comprising a breast engaging member for establishing a fluid connection between the engaging member and a nipple or alveolar duct of the subject, pressure changing means connected with said breast engaging member for changing pressure at a surface of the nipple or within the alveolar duct to induce or facilitate breast fluid expression from the nipple or alveolar duct, and a solid phase sample collection medium in fluid connection with said breast engaging member for receiving a sample of expressed breast fluid from the nipple or alveolar duct, wherein said sample comprises one or more breast disease marker(s) selected from the group consisting of a protein, a peptide, a glycoprotein, a lipid, a glycolipid, a DNA polynucleotide, an RNA polynucleotide, or a combination thereof; and    concurrent or subsequent to expression of said breast fluid, collecting the biological sample comprising the expressed mammary fluid or a component thereof that includes one or more breast disease marker(s) on or within the solid phase sample collection medium.    
   
   
       64 . The method of  claim 63 , wherein said one or more breast disease marker(s) is/are selected from the group consisting of Her 2, Ki67 Growth Factor, Cyclin B1, Cyclin D1, Proliferating Cell Nuclear Antigen, Transforming Growth Factor a, Tissue Plasminogen Activator, Insulin Growth Factor Receptors, Collagenase Type IV, Laminins, Laminin Receptor, Integrins, p53, rb, nm23, ras, c-myc, c-myb, Heat Shock Proteins, Prolactin, Neuron-Specific Enolase, IR-14, KA 1, KA 14, Alpha-Lactalbumin, Actin, CEA, HMFG, MCA, PSA, Vasopressin, Cathepsin D, PGE2, pS2; IL-10, S-100 protein; Vimentin; Epithelial Membrane Antigen, bcl-2, CA15-3, CA 19-9, Tn Antigen, Alpha-lactalbumin, LASA, Gal-GalNAC, GCDFP-15, Le(y)-Related Carbohydrate Antigen, CA 125, uPA, uPA related antigens and complexes, uPA Receptor, PAl-I and PA1-2, Beta-glucuronidase, CD31, CD44 splice variants, blood group antigens including ABH, Lewis, and MN, and genetic lesions or altered expression levels of CCND1, EMS 1, BRCA1 and BRCA2 genes, and combinations thereof.  
   
   
       65 . The method of  claim 63 , wherein said solid phase sample collection medium is in fluid contact with said nipple or alveolar duct when the breast engaging member is applied to the breast and positive or negative pressure is generated by said pressure changing means.  
   
   
       66 . The method of  claim 63 , wherein said pressure changing means is provided by said solid phase sample collection medium which operates by wicking or capillary action in contact with the nipple or alveolar duct to facilitate expression or evacuation of the sample of expressed breast fluid there from.  
   
   
       67 . The method of  claim 63 , wherein said solid phase sample collection medium is removably placed in fluid connection with a breast pump.  
   
   
       68 . The method of  claim 63 , wherein said solid phase sample collection medium is selected from the group consisting of microscopic glass slides, capillary tubes, collection tubes, columns, micro-columns, wells, plates, membranes, filters, resins, inorganic matrices, beads, resins, particulate chromatographic media, plastic microparticles, latex particles, coated tubes, coated templates, coated beads, coated matrices, and combinations thereof.  
   
   
       69 . The method of  claim 63 , wherein said biological sample is selected from the group consisting of whole mammary fluid, whole cells, cell fragments, cell membranes, proteins, glycoproteins, peptides, nucleotide components of mammary fluid, or a combination thereof.  
   
   
       70 . The method of  claim 63 , wherein said pressure changing means functions to induce or facilitate expulsion of breast fluid from the nipple or alveolar duct independently of, or in conjunction with, positive pressure generated within the alveolar duct of the breast by spontaneous or oxytocin-induced contraction, or manual compression, of the alveolar duct.  
   
   
       71 . A method for determining a presence or quantity of a breast disease marker in a biological sample of mammary fluid, comprising the steps of: 
 applying a sample collection device for collecting said biological sample from said subject comprising a breast engaging member for establishing a fluid connection between the engaging member and a nipple or alveolar duct of the subject, pressure changing means connected with said breast engaging member for changing pressure at a surface of the nipple or within the alveolar duct to induce or facilitate breast fluid expression from the nipple or alveolar duct, and a solid phase sample collection medium in fluid connection with said breast engaging member for receiving a sample of expressed breast fluid from the nipple or alveolar duct;    concurrent or subsequent to expression of said breast fluid, collecting the biological sample comprising the expressed mammary fluid or a component thereof that includes one or more breast disease marker(s) on or within the solid phase sample collection medium; and    detecting a presence or quantity of said breast disease marker in said sample.    
   
   
       72 . The method of  claim 71 , wherein said breast disease marker is a breast cancer marker.  
   
   
       73 . The method of  claim 71 , wherein said breast disease marker is from the group consisting of Her 2, Ki67 Growth Factor, Cyclin B1, Cyclin D1, Proliferating Cell Nuclear Antigen, Transforming Growth Factor a, Tissue Plasminogen Activator, Insulin Growth Factor Receptors, Collagenase Type IV, Laminins, Laminin Receptor, Integrins, p53, rb, nm23, ras, c-myc, c-myb, Heat Shock Proteins, Prolactin, Neuron-Specific Enolase, IR-14, KA 1, KA 14, Alpha-Lactalbumin, Actin, CEA, HMFG, MCA, PSA, Vasopressin, Cathepsin D, PGE2, pS2; IL-10, S-100 protein; Vimentin; Epithelial Membrane Antigen, bcl-2, CA15-3, CA 19-9, Tn Antigen, Alpha-lactalbumin, LASA, Gal-GalNAC, GCDFP-15, Le(y)-Related Carbohydrate Antigen, CA 125, uPA, uPA related antigens and complexes, uPA Receptor, PA1-1 and PA1-2, Beta-glucuronidase, CD31, CD44 splice variants, blood group antigens including ABH, Lewis, and MN, and genetic lesions or altered expression levels of CCND1, EMS1, BRCA1 and BRCA2 genes, and combinations thereof  
   
   
       74 . The method of  claim 71 , wherein said solid phase sample collection medium is selected from the group consisting of microscopic glass slides, capillary tubes, collection tubes, columns, micro-columns, wells, plates, membranes, filters, resins, inorganic matrices, beads, resins, particulate chromatographic media, plastic microparticles, latex particles, coated tubes, coated templates, coated beads, coated matrices, and combinations thereof.