Kamlesh Gidwani1, Kaisa Huhtinen2, Henna Kekki3, Sandra van Vliet4, Johanna Hynninen5, Niina Koivuviita6, Antti Perheentupa5, Matti Poutanen7, Annika Auranen8, Seija Grenman5, Urpo Lamminmäki3, Olli Carpen9, Yvette van Kooyk4, Kim Pettersson3. 1. Department of Biochemistry/Biotechnology, University of Turku, Turku, Finland; kamgid@utu.fi. 2. Department of Pathology, Medicity research laboratories, University of Turku and Turku University Hospital, Turku, Finland; 3. Department of Biochemistry/Biotechnology, University of Turku, Turku, Finland; 4. Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands; 5. Department of Obstetrics and Gynecology, University of Turku and Turku University Hospital, Turku, Finland; 6. Department of Medicine, University of Turku and Turku University Hospital, Turku, Finland; 7. Department of Physiology, Institute of Biomedicine, and Turku Center for Disease Modeling, University of Turku, Finland; 8. Department of Obstetrics and Gynecology, University of Turku and Turku University Hospital, Turku, Finland; Department of Obstetrics and Gynecology, Tampere University Hospital, Tampere, Finland; 9. Department of Pathology, Medicity research laboratories, University of Turku and Turku University Hospital, Turku, Finland; Department of Pathology, University of Helsinki and Helsinki University Hospital, Helsinki, Finland.
Abstract
BACKGROUND: Measurement of serum cancer antigen 125 (CA125) is the standard approach for epithelial ovarian cancer (EOC) diagnostics and follow-up. However, the clinical specificity is not optimal because increased values are also detected in healthy controls and in benign diseases. CA125 is known to be differentially glycosylated in EOC, potentially offering a way to construct CA125 assays with improved cancer specificity. Our goal was to identify carbohydrate-reactive lectins for discriminating between CA125 originating from EOC and noncancerous sources. METHODS: CA125 from the OVCAR-3 cancer cell line, placental homogenate, and ascites fluid from patients with cirrhosis were captured on anti-CA125 antibody immobilized on microtitration wells. A panel of lectins, each coated onto fluorescent europium-chelate-doped 97-nm nanoparticles (Eu(+3)-NPs), was tested for detection of the immobilized CA125. Serum samples from high-grade serous EOC or patients with endometriosis and healthy controls were analyzed. RESULTS: By using macrophage galactose-type lectin (MGL)-coated Eu(+3)-NPs, an analytically sensitive CA125 assay (CA125(MGL)) was achieved that specifically recognized the CA125 isoform produced by EOC, whereas the recognition of CA125 from nonmalignant conditions was reduced. Serum CA125(MGL) measurement better discriminated patients with EOC from endometriosis compared to conventional immunoassay. The discrimination was particularly improved for marginally increased CA125 values and for earlier detection of EOC progression. CONCLUSIONS: The new CA125(MGL) assay concept could help reduce the false-positive rates of conventional CA125 immunoassays. The improved analytical specificity of this test approach is dependent on a discriminating lectin immobilized in large numbers on Eu(+3)-NPs, providing both an avidity effect and signal amplification.
BACKGROUND: Measurement of serum cancer antigen 125 (CA125) is the standard approach for epithelial ovarian cancer (EOC) diagnostics and follow-up. However, the clinical specificity is not optimal because increased values are also detected in healthy controls and in benign diseases. CA125 is known to be differentially glycosylated in EOC, potentially offering a way to construct CA125 assays with improved cancer specificity. Our goal was to identify carbohydrate-reactive lectins for discriminating between CA125 originating from EOC and noncancerous sources. METHODS:CA125 from the OVCAR-3 cancer cell line, placental homogenate, and ascites fluid from patients with cirrhosis were captured on anti-CA125 antibody immobilized on microtitration wells. A panel of lectins, each coated onto fluorescent europium-chelate-doped 97-nm nanoparticles (Eu(+3)-NPs), was tested for detection of the immobilized CA125. Serum samples from high-grade serous EOC or patients with endometriosis and healthy controls were analyzed. RESULTS: By using macrophage galactose-type lectin (MGL)-coated Eu(+3)-NPs, an analytically sensitive CA125 assay (CA125(MGL)) was achieved that specifically recognized the CA125 isoform produced by EOC, whereas the recognition of CA125 from nonmalignant conditions was reduced. Serum CA125(MGL) measurement better discriminated patients with EOC from endometriosis compared to conventional immunoassay. The discrimination was particularly improved for marginally increased CA125 values and for earlier detection of EOC progression. CONCLUSIONS: The new CA125(MGL) assay concept could help reduce the false-positive rates of conventional CA125 immunoassays. The improved analytical specificity of this test approach is dependent on a discriminating lectin immobilized in large numbers on Eu(+3)-NPs, providing both an avidity effect and signal amplification.
Authors: Shruti Jain; Nimrah Nadeem; Benjamin Ulfenborg; Maria Mäkelä; Shamima Afrin Ruma; Joonas Terävä; Kaisa Huhtinen; Janne Leivo; Björg Kristjansdottir; Kim Pettersson; Karin Sundfeldt; Kamlesh Gidwani Journal: Int J Cancer Date: 2022-05-25 Impact factor: 7.316