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Serum/Plasma Proteomics pp 263–301Cite as

Surface Profiling of Extracellular Vesicles from Plasma or Ascites Fluid Using DotScan Antibody Microarrays

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1619))

Abstract

DotScan antibody microarrays were initially developed for the extensive surface profiling of live leukemia and lymphoma cells. DotScan’s diagnostic capability was validated with an extensive clinical trial using mononuclear cells from the blood or bone marrow of leukemia or lymphoma patients. DotScan has also been used for the profiling of surface proteins on peripheral blood mononuclear cells (PBMC) from patients with HIV, liver disease, and stable and progressive B-cell chronic lymphocytic leukemia (CLL). Fluorescence multiplexing allowed the simultaneous profiling of cancer cells and leukocytes from disaggregated colorectal and melanoma tumor biopsies after capture on DotScan. In this chapter, we have used DotScan for the surface profiling of extracellular vesicles (EV) recovered from conditioned growth medium of cancer cell lines and the blood of patients with CLL. The detection of captured EV was performed by enhanced chemiluminescence (ECL) using biotinylated antibodies that recognized antigens expressed on the surface of the EV subset of interest. DotScan was also used to profile EV from the blood of healthy individuals and the ascites fluid of ovarian cancer patients. DotScan binding patterns of EV from human plasma and other body fluids may yield diagnostic or prognostic signatures for monitoring the incidence, treatment, and progression of cancers.

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References

  1. D'Souza-Schorey C, Clancy JW (2012) Tumor-derived microvesicles: shedding light on novel microenvironment modulators and prospective cancer biomarkers. Genes Dev 26(12):1287–1299

    Article  PubMed  PubMed Central  Google Scholar 

  2. Taylor DD, Gercel-Taylor C (2005) Tumour-derived exosomes and their role in cancer-associated T-cell signalling defects. Br J Cancer 92(2):305–311

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Cai J, Han Y, Ren H et al (2013) Extracellular vesicle-mediated transfer of donor genomic DNA to recipient cells is a novel mechanism for genetic influence between cells. J Mol Cell Biol 5(4):227–238

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Cocucci E, Meldolesi J (2015) Ectosomes and exosomes: shedding the confusion between extracellular vesicles. Trends Cell Biol 25(6):364–372

    Article  CAS  PubMed  Google Scholar 

  5. Shifrin DA, Beckler MD, Coffey RJ et al (2013) Extracellular vesicles: communication, coercion, and conditioning. Mol Biol Cell 24(9):1253–1259

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Camussi G, Deregibus MC, Tetta C (2013) Tumor-derived microvesicles and the cancer microenvironment. Curr Mol Med 13(1):58–67

    Article  CAS  PubMed  Google Scholar 

  7. Kucharzewska P, Belting M (2013) Emerging roles of extracellular vesicles in the adaptive response of tumour cells to microenvironmental stress. J Extracell Vesicles. doi:10.3402/jev.v2i0.20304

    PubMed  PubMed Central  Google Scholar 

  8. An T, Qin S, Xu Y et al (2015) Exosomes serve as tumour markers for personalized diagnostics owing to their important role in cancer metastasis. J Extracell Vesicles. doi:10.3402/jev.v4.27522

    PubMed  PubMed Central  Google Scholar 

  9. Tatischeff I (2015) Cell-derived extracellular vesicles open new perspectives for cancer research. Cancer Res Front 1(2):208–224

    Article  Google Scholar 

  10. Webber J, Yeung V, Clayton A (2015) Extracellular vesicles as modulators of the cancer microenvironment. Semin Cell Dev Biol 40:27–34

    Article  CAS  PubMed  Google Scholar 

  11. Fujita Y, Yoshioka Y, Ochiya T (2016) Extracellular vesicle transfer of cancer pathogenic components. Cancer Sci 107(4):385–390

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Keerthikumar S, Gangoda L, Liem M et al (2015) Proteogenomic analysis reveals exosomes are more oncogenic than ectosomes. Oncotarget 6(17):15375–15396. doi:10.18632/oncotarget.3801

    Article  PubMed  PubMed Central  Google Scholar 

  13. Xu R, Greening DW, Rai A et al (2015) Highly-purified exosomes and shed microvesicles isolated from the human colon cancer cell line LIM1863 by sequential centrifugal ultrafiltration are biochemically and functionally distinct. Methods 87:11–25

    Article  CAS  PubMed  Google Scholar 

  14. Green TM, Alpaugh ML, Barsky SH et al (2015) Breast cancer-derived extracellular vesicles: characterization and contribution to the metastatic phenotype. Biomed Res Int. doi:10.1155/2015/634865

    Google Scholar 

  15. Santiago-Dieppa DR, Steinberg J, Gonda D et al (2014) Extracellular vesicles as a platform for ‘liquid biopsy’in glioblastoma patients. Expert Rev Mol Diagn 14(7):819–825

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Mizutani K, Terazawa R, Kameyama K et al (2014) Isolation of prostate cancer-related exosomes. Anticancer Res 34(7):3419–3423

    CAS  PubMed  Google Scholar 

  17. Royo F, Zuñiga-Garcia P, Torrano V et al (2016) Transcriptomic profiling of urine extracellular vesicles reveals alterations of CDH3 in prostate cancer. Oncotarget 7(6):6835–6846

    PubMed  PubMed Central  Google Scholar 

  18. Gámez-Valero A, Lozano-Ramos SI, Bancu I et al (2015) Urinary extracellular vesicles as source of biomarkers in kidney diseases. Front Immunol 6:6. doi:10.3389/fimmu.2015.00006

    Article  PubMed  PubMed Central  Google Scholar 

  19. Runz S, Keller S, Rupp C et al (2007) Malignant ascites-derived exosomes of ovarian carcinoma patients contain CD24 and EpCAM. Gynecol Oncol 107(3):563–571

    Article  CAS  PubMed  Google Scholar 

  20. Elashoff D, Zhou H, Reiss J et al (2012) Prevalidation of salivary biomarkers for oral cancer detection. Cancer Epidemiol Biomark Prev 21(4):664–672

    Article  CAS  Google Scholar 

  21. Nicholas J (2013) A new diagnostic tool with the potential to predict tumor metastasis. J Natl Cancer Inst 105(6):371-372

    Google Scholar 

  22. Kelleher RJ, Balu-Iyer S, Loyall J et al (2015) Extracellular vesicles present in human ovarian tumor microenvironments induce a phosphatidylserine-dependent arrest in the T-cell signaling cascade. Cancer Immunol Res 3(11):1269–1278

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Momen-Heravi F, Balaj L, Alian S, et al. 2012 Alternative methods for characterization of extracellular vesicles Front Physiol 3(354.103389

    Google Scholar 

  24. Belov L, Mulligan SP, Barber N et al (1999) Analysis of human leukaemias and lymphomas using extensive immunophenotypes from an antibody microarray. Br J Haematol 135(2):184–197

    Article  Google Scholar 

  25. Ellmark P, Belov L, Huang P et al (2006) Multiplex detection of surface molecules on colorectal cancers. Proteomics 6(6):1791–1802

    Article  CAS  PubMed  Google Scholar 

  26. Kaufman KL, Belov L, Huang P et al (2010) An extended antibody microarray for surface profiling metastatic melanoma. J Immunol Methods 358(1-2):23–34

    Article  CAS  PubMed  Google Scholar 

  27. Zhou J, Belov L, Huang PY et al (2010) Surface antigen profiling of colorectal cancer using antibody microarrays with fluorescence multiplexing. J Immunol Methods 355(1-2):40–51

    Article  CAS  PubMed  Google Scholar 

  28. Zhou J, Belov L, Chapuis P et al (2015) Surface profiles of live colorectal cancer cells and tumor infiltrating lymphocytes from surgical samples correspond to prognostic categories. J Immunol Methods 416:59–68

    Article  CAS  PubMed  Google Scholar 

  29. Zhou J, Belov L, Solomon MJ et al (2011) Colorectal cancer cell surface protein profiling using an antibody microarray and fluorescence multiplexing. J Vis Exp 55:3322. doi:10.3791/3322

    Google Scholar 

  30. Wu JQ, Dyer WB, Chrisp J et al (2008) Longitudinal microarray analysis of cell surface antigens on peripheral blood mononuclear cells from HIV+ individuals on highly active antiretroviral therapy. Retrovirology 5(1):24–35. doi:10.1186/1742-4690-5-24

    Article  PubMed  PubMed Central  Google Scholar 

  31. Wu JQ, Wang B, Belov L et al (2007) Antibody microarray analysis of cell surface antigens on CD4+ and CD8+ T cells from HIV+ individuals correlates with disease stages. Retrovirology 4(1):83–95. doi:10.1186/1742-4690-4-83

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Woolfson A, Stebbing J, Tom BD et al (2005) Conservation of unique cell-surface CD antigen mosaics in HIV-1-infected individuals. Blood 106(3):1003–1007

    Article  CAS  PubMed  Google Scholar 

  33. Rahman W, Huang P, Belov L et al (2012) Analysis of human liver disease using a cluster of differentiation (CD) antibody microarray. Liver Int 32(10):1527–1534

    Article  PubMed  Google Scholar 

  34. Rahman W, Tu T, Budzinska M et al (2015) Analysis of post-liver transplant hepatitis C virus recurrence using serial cluster of differentiation antibody microarrays. Transplantation 99(9):e120–e126

    Article  CAS  PubMed  Google Scholar 

  35. Huang PY, Kohnke P, Belov L et al (2013) Profiles of surface mosaics on chronic lymphocytic leukemias distinguish stable and progressive subtypes. J Pharm Pharm Sci 16(2):231–237

    Article  PubMed  Google Scholar 

  36. Ellmark P, Högerkorp C-M, Ek S et al (2008) Phenotypic protein profiling of different B cell sub-populations using antibody CD-microarrays. Cancer Lett 265(1):98–106

    Article  CAS  PubMed  Google Scholar 

  37. Siegel R, Ma J, Zou Z et al (2014) Cancer statistics, 2014. CA Cancer J Clin 64(1):9–29

    Article  PubMed  Google Scholar 

  38. Ghosh AK, Secreto CR, Knox TR et al (2010) Circulating microvesicles in B-cell chronic lymphocytic leukemia can stimulate marrow stromal cells: implications for disease progression. Blood 115(9):1755–1764

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Chiorazzi N, Ferrarini M (2003) B cell chronic lymphocytic leukemia: lessons learned from studies of the B cell antigen receptor. Annu Rev Immunol 21(1):841–894

    Article  CAS  PubMed  Google Scholar 

  40. Seifert M, Sellmann L, Bloehdorn J et al (2012) Cellular origin and pathophysiology of chronic lymphocytic leukemia. J Exp Med 209(12):2183–2198

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Lee M (2014) Prognostic impact of epithelial cell adhesion molecule in ovarian cancer patients. J Gynecol Oncol 25(4):352–354

    Article  PubMed  PubMed Central  Google Scholar 

  42. Ahmed N, Stenvers KL (2013) Getting to know ovarian cancer ascites: opportunities for targeted therapy-based translational research. Front Oncol 3:256. doi:10.3389/fonc.2013.00256

    Article  PubMed  PubMed Central  Google Scholar 

  43. Belov L, Matic KJ, Hallal S et al (2016) Extensive surface protein profiles of extracellular vesicles from cancer cells may provide diagnostic signatures from blood samples. J Extracell Vesicles 5:25355. doi:10.3402/jev.v5.25355

    Article  PubMed  Google Scholar 

  44. Kanada M, Bachmann MH, Hardy JW et al (2015) Differential fates of biomolecules delivered to target cells via extracellular vesicles. Proc Natl Acad Sci U S A 112(12):E1433–E1442

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Lötvall J, Hill AF, Hochberg F et al (2014) Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J Extracell Vesicles 3:26913. doi:10.3402/jev.v3.26913

    Article  PubMed  Google Scholar 

  46. Crescitelli R, Lässer C, Szabo TG et al (2013) Distinct RNA profiles in subpopulations of extracellular vesicles: apoptotic bodies, microvesicles and exosomes. J Extracell Vesicles 2:20677. doi:10.3402/jev.v2i0

    Article  Google Scholar 

  47. Luo X, Fan Y, Park I-W et al (2015) Exosomes are unlikely involved in intercellular nef transfer. PLoS One 10(4):e0124436. doi:10.1371/journal.pone.0124436

    Article  PubMed  PubMed Central  Google Scholar 

  48. Oksvold MP, Kullmann A, Forfang L et al (2014) Expression of B-cell surface antigens in subpopulations of exosomes released from B-cell lymphoma cells. Clin Ther 36(6):847–862

    Article  CAS  PubMed  Google Scholar 

  49. Saunderson SC, Schuberth PC, Dunn AC et al (2008) Induction of exosome release in primary B cells stimulated via CD40 and the IL-4 receptor. J Immunol 180(12):8146–8152

    Article  CAS  PubMed  Google Scholar 

  50. Booth AM, Fang Y, Fallon JK et al (2006) Exosomes and HIV gag bud from endosome-like domains of the T cell plasma membrane. J Cell Biol 172(6):923–935

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Fang Y, Wu N, Gan X et al (2007) Higher-order oligomerization targets plasma membrane proteins and HIV gag to exosomes. PLoS Biol 5(6):e158. doi:10.1371/journal.pbio.0050158

    Article  PubMed  PubMed Central  Google Scholar 

  52. Andreu Z, Yáñez-Mó M (2014) Tetraspanins in extracellular vesicle formation and function. Front Immunol 5:442. doi:10.3389/fimmu.2014.00442

    Article  PubMed  PubMed Central  Google Scholar 

  53. Thery C, Amigorena S, Raposo G et al (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol Chapter 3:Unit 22. doi:10.1002/0471143030.cb0322s30

    PubMed  Google Scholar 

  54. Logozzi M, De Milito A, Lugini L et al (2009) High levels of exosomes expressing CD63 and caveolin-1 in plasma of melanoma patients. PLoS One 4(4):e5219. doi:10.1371/journal.pone.0005219

    Article  PubMed  PubMed Central  Google Scholar 

  55. Colino J, Snapper CM (2007) Dendritic cell-derived exosomes express a Streptococcus Pneumoniae capsular polysaccharide type 14 cross-reactive antigen that induces protective immunoglobulin responses against pneumococcal infection in mice. Infect Immun 75(1):220–230

    Article  CAS  PubMed  Google Scholar 

  56. Lai RC, Tan SS, Teh BJ et al (2012) Proteolytic potential of the MSC exosome proteome: implications for an exosome-mediated delivery of therapeutic proteasome. Int J Proteomics. doi:10.1155/2012/971907

    Google Scholar 

  57. Kalra H, Adda CG, Liem M et al (2013) Comparative proteomics evaluation of plasma exosome isolation techniques and assessment of the stability of exosomes in normal human blood plasma. Proteomics 13(22):3354–3364

    Article  CAS  PubMed  Google Scholar 

  58. Ellmark P, Woolfson A, Belov L et al (2008) The applicability of a cluster of differentiation monoclonal antibody microarray to the diagnosis of human disease. Methods Mol Biol 439:199–209. doi:10.1007/978-1-59745-188-8_14

    Article  CAS  PubMed  Google Scholar 

  59. Brown A, Lattimore JD, McGrady M et al (2008) Stable and unstable angina: identifying novel markers on circulating leukocytes. Proteomics Clin Appl 2(1):90–98

    Article  CAS  PubMed  Google Scholar 

  60. Stacchini A, Aragno M, Vallario A et al (1999) MEC1 and MEC2: two new cell lines derived from B-chronic lymphocytic leukaemia in prolymphocytoid transformation. Leuk Res 23(2):127–136

    Article  CAS  PubMed  Google Scholar 

  61. Hamelinck D, Zhou H, Li L et al (2005) Optimized normalization for antibody microarrays and application to serum-protein profiling. Mol Cell Proteomics 4(6):773–784

    Article  CAS  PubMed  Google Scholar 

  62. Zhou J, Belov L, Armstrong N et al (2013) Antibody microarrays and multiplexing. In: Wang X (ed) Bioinformatics of human proteomics. Springer, New York, NY, pp 331–359. doi:10.1007/978-94-007-5811-7_15

    Chapter  Google Scholar 

  63. Belov L, de la Vega O, dos Remedios CG et al (2001) Immunophenotyping of leukemias using a cluster of differentiation antibody microarray. Cancer Res 61(11):4483–4489

    CAS  PubMed  Google Scholar 

  64. Austin J, Holway AH (2011) Contact printing of protein microarrays. Methods Mol Biol 785:379–394. doi:10.1007/978-1-61779-286-1_25

    Article  CAS  PubMed  Google Scholar 

  65. Seurynck-Servoss SL, White AM, Baird CL et al (2007) Evaluation of surface chemistries for antibody microarrays. Anal Biochem 371(1):105–115

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Li J, Lee Y, Johansson HJ et al (2015) Serum-free culture alters the quantity and protein composition of neuroblastoma-derived extracellular vesicles. J Extracell Vesicles 4:26883. doi:10.3402/jev.v4.26883

    Article  PubMed  Google Scholar 

  67. Shelke GV, Lässer C, Gho YS et al (2014) Importance of exosome depletion protocols to eliminate functional and RNA-containing extracellular vesicles from fetal bovine serum. J Extracell Vesicles 3:24783. doi:10.3402/jev.v3

    Article  Google Scholar 

  68. György B, Pálóczi K, Kovács A et al (2014) Improved circulating microparticle analysis in acid-citrate dextrose (ACD) anticoagulant tube. Thromb Res 133(2):285–292

    Article  PubMed  Google Scholar 

  69. Hallek M, Cheson BD, Catovsky D et al (2008) Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the international workshop on chronic lymphocytic leukemia updating the National Cancer Institute–working group 1996 guidelines. Blood 111(12):5446–5456

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Adcock DM, Kressin DC, Marlar RA et al (1997) Effect of 3.2% vs 3.8% sodium citrate concentration on routine coagulation testing. Am J Clin Pathol 107(1):105–110

    Article  CAS  PubMed  Google Scholar 

  71. Sedlmayr P, Leitner V, Pilz S et al (2001) Species-specific blocking of fc-receptors in indirect immunofluorescence assays. Lab Hematol 7:81–84

    CAS  Google Scholar 

  72. Gardiner C, Ferreira YJ, Dragovic RA (2013) Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis. J Extracell Vesicles 2:19671. doi:10.3402/jev.v2i0.19671

    Article  Google Scholar 

  73. Trummer A, De Rop C, Tiede A et al (2008) Isotype controls in phenotyping and quantification of microparticles: a major source of error and how to evade it. Thromb Res 122(5):691–700

    Article  CAS  PubMed  Google Scholar 

  74. Amadori A, Zamarchi R, De Silvestro G et al (1995) Genetic control of the CD4/CD8 T-cell ratio in humans. Nat Med 1(12):1279–1283

    Article  CAS  PubMed  Google Scholar 

  75. Gibbings DJ, Marcet-Palacios M, Sekar Y et al (2007) CD8α is expressed by human monocytes and enhances FcγR-dependent responses. BMC Immunol 8(1):12. doi:10.1186/1471-2172-8-12

    Article  PubMed  PubMed Central  Google Scholar 

  76. Campbell JP, Guy K, Cosgrove C et al (2008) Total lymphocyte CD8 expression is not a reliable marker of cytotoxic T-cell populations in human peripheral blood following an acute bout of high-intensity exercise. Brain Behav Immun 22(3):375–380

    Article  CAS  PubMed  Google Scholar 

  77. van Velzen JF, Laros-van Gorkom BA, Pop GA (2012) Multicolor flow cytometry for evaluation of platelet surface antigens and activation markers. Thromb Res 130(1):92–98

    Article  PubMed  Google Scholar 

  78. Kubagawa H, Oka S, Kubagawa Y et al (2009) Identity of the elusive IgM fc receptor (FcμR) in humans. J Exp Med 206(12):2779–2793

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  79. Varki A, Gagneux P (2012) Multifarious roles of sialic acids in immunity. Ann N Y Acad Sci 1253(1):16–36

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Heijnen HF, Schiel AE, Fijnheer R et al (1999) Activated platelets release two types of membrane vesicles: microvesicles by surface shedding and exosomes derived from exocytosis of multivesicular bodies and alpha-granules. Blood 94(11):3791–3799

    CAS  PubMed  Google Scholar 

  81. Colucci M, Stöckmann H, Butera A et al (2015) Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. J Immunol 194(1):151–157

    Article  CAS  PubMed  Google Scholar 

  82. Arnold JN, Wormald MR, Suter DM et al (2005) Human serum IgM glycosylation: identification of glycoforms that can bind to mannan-binding lectin. J Biol Chem 280(32):29080–29087

    Article  CAS  PubMed  Google Scholar 

  83. Creasey AM, Staalsoe T, Raza A et al (2003) Nonspecific immunoglobulin M binding and chondroitin sulfate a binding are linked phenotypes of plasmodium falciparum isolates implicated in malaria during pregnancy. Infect Immun 71(8):4767–4771

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Molica S, Mauro FR, Giannarelli D et al (2011) Differentiating chronic lymphocytic leukemia from monoclonal B-lymphocytosis according to clinical outcome: on behalf of the GIMEMA chronic lymphoproliferative diseases working group. Haematologica 96(2):277–283

    Article  PubMed  Google Scholar 

  85. Shanafelt T (2013) Treatment of older patients with chronic lymphocytic leukemia: key questions and current answers. ASH Educ Program Book 2013(1):158–167

    Google Scholar 

  86. Dearden C (2008) Disease-specific complications of chronic lymphocytic leukemia. ASH Educ Program Book 2008(1):450–456

    Google Scholar 

  87. Pugholm LH, Bæk R, Søndergaard EKL et al (2016) Phenotyping of leukocytes and leukocyte-derived extracellular vesicles. J Immunol Res 2016:6391264. doi:10.1155/2016/6391264

    Article  PubMed  PubMed Central  Google Scholar 

  88. Mathivanan S, Lim JW, Tauro BJ et al (2010) Proteomics analysis of A33 immunoaffinity-purified exosomes released from the human colon tumor cell line LIM1215 reveals a tissue-specific protein signature. Mol Cell Proteomics 9(2):197–208

    Article  CAS  PubMed  Google Scholar 

  89. Newland RC, Chapuis PH, Pheils MT et al (1981) The relationship of survival to staging and grading of colorectal carcinoma: a prospective study of 503 cases. Cancer 47(6):1424–1429

    Article  CAS  PubMed  Google Scholar 

  90. Davis NC, Evans EB, Cohen JR et al (1984) Staging of colorectal cancer. The Australian clinico-pathological staging (ACPS) system compared with Dukes’ system. Dis Colon Rectum 27(11):707–713

    Article  CAS  PubMed  Google Scholar 

  91. Tarantino I, Warschkow R, Worni M et al (2012) Elevated preoperative CEA is associated with worse survival in stage I–III rectal cancer patients. Br J Cancer 107(2):266–274

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Di Matteo P, Arrigoni GL, Alberici L et al (2011) Enhanced expression of CD13 in vessels of inflammatory and neoplastic tissues. J Histochem Cytochem 59(1):47–59

    Article  PubMed  PubMed Central  Google Scholar 

  93. Ruibal MA (1991) CEA serum levels in non-neoplastic disease. Int J Biol Markers 7(3):160–166

    Google Scholar 

  94. Tesselaar ME, Romijn FP, Van Der Linden IK et al (2007) Microparticle-associated tissue factor activity: a link between cancer and thrombosis? J Thromb Haemost 5(3):520–527

    Article  CAS  PubMed  Google Scholar 

  95. Moreno A, Lucena C, Lopez A et al (2002) Immunohistochemical analysis of ß3 integrin (CD61): expression in pig tissues and human tumors. Histol Histopathol 17:347–352

    CAS  Google Scholar 

  96. Alomari M (2014) Proteomic characterisation of chronic lymphocytic leukaemia cells treated with rituximab. Thesis- http://hdl.handle.net/2123/12407

  97. Golay J, Lazzari M, Facchinetti V et al (2001) CD20 levels determine the in vitro susceptibility to rituximab and complement of B-cell chronic lymphocytic leukemia: further regulation by CD55 and CD59. Blood 98(12):3383–3389

    Article  CAS  PubMed  Google Scholar 

  98. Cantor JM, Ginsberg MH (2012) CD98 at the crossroads of adaptive immunity and cancer. J Cell Sci 125(6):1373–1382

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

We wish to thank Dr. Pauline Huang (School of Life and Environmental Sciences, University of Sydney, Australia) for making the DotScan antibody microarrays and A/Prof. Philip Beale (Concord Repatriation General Hospital, Concord, NSW, Australia) for supplying blood samples from CRC patients.

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Authors and Affiliations

  1. School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, 2006, Australia

    Larissa Belov, Susannah Hallal, Kieran Matic, Jerry Zhou, Sandra Wissmueller, Stephen P. Mulligan, O. Giles Best & Richard I. Christopherson

  2. Fiona Elsey Cancer Research Institute, Ballarat, VIC, 3350, Australia

    Nuzhat Ahmed

  3. Federation University, Ballarat, VIC, 3355, Australia

    Nuzhat Ahmed

  4. Department of Obstetrics & Gynaecology, Women’s Cancer Research Centre, Royal Women’s Hospital, Parkville, VIC, 3052, Australia

    Sumaiya Tanjil

  5. Kolling Institute of Medical Research, Royal North Shore Hospital, St. Leonards, NSW, 2065, Australia

    Stephen P. Mulligan & O. Giles Best

  6. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC, 3086, Australia

    Richard J. Simpson

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  1. Larissa Belov
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  9. O. Giles Best
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  10. Richard J. Simpson
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  11. Richard I. Christopherson
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Correspondence to Larissa Belov .

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Editors and Affiliations

  1. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria, Australia

    David W. Greening

  2. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria, Australia

    Richard J. Simpson

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Belov, L. et al. (2017). Surface Profiling of Extracellular Vesicles from Plasma or Ascites Fluid Using DotScan Antibody Microarrays. In: Greening, D., Simpson, R. (eds) Serum/Plasma Proteomics. Methods in Molecular Biology, vol 1619. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7057-5_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7057-5_20

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