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  • American Association for Cancer Research (AACR)  (8)
  • 1
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 77, No. 13_Supplement ( 2017-07-01), p. 1721-1721
    Abstract: Background: Circulating tumor cells (CTCs) are tumor cells shed from primary tumor and circulate in the peripheral blood. CTCs, as a surrogate of distant metastasis, can be potentially useful in diagnosis and monitoring therapeutic effects in malignant tumors. Among a variety of systems for detection of CTCs, the “Cellsearch” is the only approved system for clinical use. However, EpCAM-negative tumor cells, such as those originating from non-epithelial cells and those undergoing epithelial-mesenchymal transition (EMT) cannot be captured with the “CellSearch” that is an EpCAM-based isolation system. Therefore, we have developed a novel polymeric microfluidic device (“Universal” CTC-chip) that can capture CTCs with or without EpCAM expression (AACR 2015). In the present study, we examined CTCs-detection performance of the CTC-chip in patients with thoracic malignant tumors (lung cancer [LC] as an “EpCAM-positive” tumor and malignant pleural mesothelioma [MPM] as an “EpCAM-negative” tumor) in comparison with that of the CellSearch. Methods: Peripheral blood sampled from each patient was divided and subjected to quantitative evaluation of CTCs with the CTC-chip as well as with the “CellSearch”. The CTC-chip, coated with an anti-EpCAM antibody, was used to capture CTCs in the blood samples (n=19) from lung cancer patients. To capture CTCs in the samples (n=11) from MPM patients, the CTC-chip was coated with an antibody against podoplanin that is expressed on the mesothelioma. After immuno-staining for cytokeratin and CD45 on the chip, a captured cell containing Hoechst-positive nucleus and cytokeratin-positive/ CD45-negative cytoplasm was judged as a CTC. The CTC-count for each sample was represented as the number per 7.5mL of the blood. Results: The median CTC-count detected with the CTC-chip in LC was 50 (range, 0-270), which was significantly higher than that (the median CTC-count, 0; range, 0-47) with the CellSearch (p & lt;0.01). In the peripheral blood sampled from MPM patients, CTC was detected in only one patient using the CellSearch, but was detected in all 11 patients with the median CTC-count of 144 (range 0-470). Conclusion: The “universal” CTC-chip achieved higher performance in detection of CTCs of thoracic malignant tumors as compared with the CellSearch. The updated data will be presented at the AACR annual meeting 2017. Citation Format: Kazue Yoneda, Taiji Kuwata, Yasuhiro Chikaishi, Kenichi Kobayashi, Sakiko Yura, Hiroki Matsumiya, Masatoshi Kanayama, Akihiro Taira, Yusuke Nabe, Shinji Shinohara, Masaru Takenaka, Soichi Oka, Ayako Hirai, Yuko Tashima, Naoko Imanishi, Koji Kuroda, Fumihiro Tanaka. Detection of CTCs in thoracic malignant tumors with "universal" CTC-chip [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1721. doi:10.1158/1538-7445.AM2017-1721
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2017
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  • 2
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    Online Resource
    American Association for Cancer Research (AACR) ; 2016
    In:  Cancer Research Vol. 76, No. 14_Supplement ( 2016-07-15), p. 3963-3963
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 76, No. 14_Supplement ( 2016-07-15), p. 3963-3963
    Abstract: Backgrounds: Circulating tumor cells (CTCs), as a surrogate of distant metastasis, can be potentially useful in early diagnosis and monitoring therapeutic effects for patients with malignant tumors. Among a variety of systems for detection of CTCs, an epithelial cell adhesion molecule (EpCAM)-based immuno-magnetic separation system “CellSearch” is the only system approved for clinical use. Our previous study showed that CTC detected by CellSearch was useful in the diagnosis or the prognosis of epithelioid-type malignant pleural mesothelioma, however, the sensitivity was insufficient. Because EpCAM-negative tumor cells, such as those originating from non-epithelial cells and those undergoing epithelial-mesenchymal transition, cannot be captured using the “CellSearch”. Therefore, we have developed a novel polymeric microfluidic device system “universal CTC-chip” that can capture a variety of CTCs with or without EpCAM expression (AACR 2015). In the present study, we assessed its sensitivity for capturing mesothelioma cells and compare with CellSearch. Methods: PC-9, human lung cancer cell line and ACC-MESO-4, human mesothelioma cell line were used in this study. Antibodies were coated on the chip in two steps, first with anti-mouse IgG antibody (“base-antibody”) and then as a “capture-antibody”, either mouse anti-human EpCAM antibody to capture EpCAM-positive cells (“EpCAM chip”) or mouse anti-human podoplanin antibody to capture podoplanin-positive mesothelioma cells (“podoplanin chip”). For sensitivity test, each cell line was suspended as concentration 500, 100, 50, 10cells/mL in phosphate buffered saline (PBS) or in blood. After 1ml of sample was flowed, we counted cells and calculated capture efficiency (number of capture cells /flowed cell). For comparison with CellSearch (CS), each cell line was suspended as concentration 50 or 10 cells /mL in blood and calculated detection efficiency (number of detected cells/ suspended cell). Results: In sensitivity test, capture efficiency of PC-9 or ACC-MESO-4 was 101.1%, 90.7% (500 cells/mL), 94.8%, 95.9% (100 cells/mL), 104.7%, 97.9% (50 cells/mL), 114.3%, 113.75% (10 cells/mL), respectively in PBS. In the blood sample, capture efficiency of PC-9 or MESO-4 was 99.2%, 50% (50 cells/mL), 115.5%, 80.55% (10 cells/mL), respectively. On the other hand, detection efficiency of PC-9 (EpCAM chip, CS) was 110%, 121.3% at 50 cells/mL, 195%, 126% at 10 cells/mL in blood. In case of MESO-4, detection efficiency (podoplanin chip, CS) was 55%, 1.1% at 50 cells/mL, 70%, 0% at 10 cells/mL in blood. Conclusion: The novel “universal CTC-chip” can be a promising CTC detection system. Citation Format: Kazue Yoneda, Yasuhiro Chikaishi, Eri Kawashima, Takashi Ohnaga, Fumihiro Tanaka. Capture of EpCAM-negative mesothelioma cells with a “universal CTC-chip”: Sensitivity test and comparison with EpCAM-based standard method. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3963.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2016
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  • 3
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    American Association for Cancer Research (AACR) ; 2015
    In:  Cancer Research Vol. 75, No. 15_Supplement ( 2015-08-01), p. 380-380
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 75, No. 15_Supplement ( 2015-08-01), p. 380-380
    Abstract: Background: Circulating tumor cell (CTC) can be a potentially useful clinical marker in early diagnosis and monitoring therapeutic effects for patients with malignant tumors. Our previous study showed that CTC detected by an EpCAM (epithelial cell adhesion molecule) based immuno-magnetic separation system “CellSearch” was useful in the diagnosis of malignant pleural mesothelioma (MPM), and also useful in the prognosis of epithelioid type MPM (Yoneda K, et al. Ann Surg Oncol. 2013). However, EpCAM-negative tumor cells cannot be principally captured with the system. Therefore, we have developed EpCAM-independent “Universal CTC-Chip System”, and assessed its capture capability. Methods: PC-9 (lung cancer cell line) was employed as EpCAM-positive cells, ACC-MESO-1 and ACC-MESO-4 (mesothelioma cell lines) were employed as EpCAM-negative cells. The expression of cell surface antigen was confirmed by flow cytometry. A microfluidic devices made of resin was coated in two steps with bridging antibody and capture antibody. It is possible to use any antibody to capture. In this study, we use anti-EpCAM antibody, anti-podoplain antibody, anti-mesothelin antibody, and isotype control. First, we verified capture capability of this system using samples which were spiked tumor cells labeled with CFSE in PBS containing 5% BSA. Then, we measured in the same manner by adding the cells to the blood of healthy donor, and calculated capture rate. Results: The expression analysis of cell surface antigens, PC-9 was EpCAM-positive, podoplanin, mesothelin were negative. On the other hand, MESO-1, 4 was EpCAM negative, podoplanin, mesothelin were positive. The capture efficiency in PBS containing 5% BSA with each cell using anti-EpCAM antibody, anti-podoplanin antibody and anti-mesothelin were PC-9: 101.1% / 2.3% / 2.9%, MESO-1: 3.5% / 52.7% / 4.3%, MESO-4: 3.0% / 78.3% / 5.4%, respectively. In the study of blood, each of the capture rate using anti-EpCAM antibody and anti-podoplanin antibody were PC-9: 88.0/ 6.9%, MESO-1: 4.0%/ 12.7%, MESO-4: 2.2%/ 38.4%, respectively. Conclusions: The capture efficiency using this device depends on the expression intensity of cell surface antigens. In experiments with PBS samples, it was possible to capture targeting specific cell surface antigen. Mesothelioma cells in the blood ware captured with combination of this CTC-Chip system and anti-podoplanin antibody. Citation Format: Kazue Yoneda, Yasuhiro Chikaishi, Eri Kawashima, Tomoko So, Hidetaka Uramoto, Takashi Ohnaga, Fumihiro Tanaka. Capture of EpCAM-negative circulating tumor cells (CTCs) with a “Universal CTC-Chip”. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 380. doi:10.1158/1538-7445.AM2015-380
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2015
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  • 4
    In: Clinical Cancer Research, American Association for Cancer Research (AACR), Vol. 15, No. 22 ( 2009-11-15), p. 6980-6986
    Abstract: Purpose: To investigate the diagnostic performance of circulating tumor cells (CTC) in discrimination between primary lung cancer and nonmalignant diseases as well as in prediction of distant metastasis. Patients and Methods: We prospectively evaluated CTCs in 7.5-mL samples of peripheral blood sampled from patients with a suspicion or a diagnosis of primary lung cancer. A semiautomated system was used to capture CTCs with an antibody against epithelial cell adhesion molecule. Results: Of 150 eligible patients, 25 were finally diagnosed as having nonmalignant disease, and 125 were diagnosed as having primary lung cancer with (n = 31) or without (n = 94) distant metastasis. CTCs were detected in 30.6 of lung cancer patients and in 12.0 of nonmalignant patients. CTC count was significantly higher in lung cancer patients than in nonmalignant patients, but a receiver operating characteristic (ROC) curve analysis showed an insufficient capability of the CTC test in discrimination between lung cancer and nonmalignant diseases with an area under ROC curve of 0.598 (95 confidence interval, 0.488-0.708; P = 0.122). Among lung cancer patients, CTC count significantly increased along with tumor progression, especially with development of distant metastasis. The area under ROC curve for CTC count in prediction of distant metastasis was 0.783 (95 confidence interval, 0.679-0.886; P & lt; 0.001). When patients with one or more CTCs were judged as having metastatic disease, sensitivity and specificity of the CTC test were 71.0 and 83.0, respectively. Conclusions: CTC is a useful surrogate marker of distant metastasis in primary lung cancer. (Clin Cancer Res 2009;15(22):69806)
    Type of Medium: Online Resource
    ISSN: 1078-0432 , 1557-3265
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2009
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  • 5
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    Online Resource
    American Association for Cancer Research (AACR) ; 2016
    In:  Cancer Research Vol. 76, No. 7_Supplement ( 2016-04-01), p. A51-A51
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 76, No. 7_Supplement ( 2016-04-01), p. A51-A51
    Abstract: Background: Circulating tumor cells (CTCs) are tumor cells shed from primary tumor and circulate in the peripheral blood. CTCs, as a surrogate of distant metastasis, can be potentially useful in diagnosis and monitoring therapeutic effects in malignant tumors. Among a variety of systems for detection of CTCs, the Cellsearch is the only system approved for clinical use. However, EpCAM-negative tumor cells, such as those originating from non-epithelial cells and those undergoing epithelial-mesenchymal transition, cannot be captured using the CellSearch that is an EpCAM-based immuno-magnetic separation system. Therefore, we have developed a novel polymeric microfluidic device system Universal CTC-chip that can capture a variety of CTCs with or without EpCAM expression (AACR 2015). In the present study, we further optimized its performance of capturing CTCs, especially EpCAM-negative CTCs. Methods: The CTC-chip was used after two-step coating, first with a goat anti-mouse IgG antibody as base-antibody and then with a capture-antibody, either a mouse anti-EpCAM antibody to capture EpCAM-positive cells (EpCAM-chip) or a mouse anti-podoplanin antibody to capture podoplanin-positive cells (podoplanin chip). An EpCAM-positive cells (PC-9: lung cancer cell line) or an EpCAM-negative and podoplanin-positive cells (ACC-MESO-4: mesothelioma cell line) were suspended in phosphate-buffered saline (PBS) or in blood, and tumor cells were captured using the CTC-chip coated with a base-antibody and a capture-antibody at different concentrations. Results: EpCAM-positive PC-9 cells were effectively captured using the EpCAM-chip coated with the base-antibody and the capture-antibody even at a lower concentration of 20μg/mL (capture rates, 101% for PBS-suspension and 88% for blood-suspension, respectively). EpCAM-negative MESO-4 cells were captured using the podoplanin-chip coated with antibodies at the lower concentration (20μg/mL), but the capture rates were lower especially for blood-suspension (78% for PBS-suspension and 38% for blood suspension, respectively). When the base-antibody was used at a higher concentration (200 or 500μg/mL) for coating the podoplanin-chip, capture efficiencies were improved for PBS-suspension (capture rates, 91% for 200μg/mL and 101% for 500μg/mL) and for blood-suspension (capture rates, 84% for 200μg/mL and 81% for 500μg/mL); even when the capture-antibody was used at a higher concentration (200μg/mL) in combination with the highest concentration of the base-antibody (500μg/mL), capture efficiencies were not further improved (capture rates, 97% for PBS-suspension and 82% for blood-suspension, respectively). Conclusion: The efficiency in capturing CTCs using the Universal CTC-chip was improved by optimizing conditions of coating the chip. Citation Format: Kazue Yoneda, Yasuhiro Chikaishi, Eri Kawashima, Takashi Ohnaga, Fumihiro Tanaka. Improved efficacy in capturing EpCAM-negative tumor cells using a universal CTC-chip. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A51.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2016
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  • 6
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 79, No. 13_Supplement ( 2019-07-01), p. 3149-3149
    Abstract: Aim: To investigate the prognostic impact of programmed death-ligand 1 (PD-L1) expression on tumor cells in correlation with neutrophil-to-lymphocyte ratio (NLR) in early-stage squamous cell carcinoma of the lung, as PD-L1 is a potent regulator of cancer immunity and NLR is a potential surrogate of immune status. Patients and methods: Eighty-three patients with completely resected pathologic stage I lung squamous cell carcinoma were retrospectively reviewed. Tumoral PD-L1 expression was evaluated with immunohistochemistry, and its prognostic impact was analyzed in correlation with NLR. Results: Forty-three patients (51.8%) had tumor with positive PD-L1 expression (percentage of tumor cells expressing PD-L1, ≥1%). There was no significant correlation between PD-L1 expression and NLR. PD-L1-positivity failed to provide a significant prognostic impact (overall survival [OS] rate at 5 years, 70.1% in PD-L1-negative patients versus 53.0% in PD-L1-positive patients; P=0.117). Among NLR-low ( & lt;2.2) patients, however, PD-L1-positivity was significantly correlated with a poor prognosis (OS rate at 5 years, 86.0% in PD-L1-negative patients versus 46.1% in PD-L1-positive patients; P=0.020). In contrast, among NLR-high (≥2.2) patients, PD-L1-positivity provided no prognostic impact (P=0.680). When NLR status and tumoral PD-L1 status were combined, “NLR-low and tumoral PD-L1-negative” was a significant and independent factor to predict a favorable recurrence-free survival (hazard ratio, 0.237 [95% confidence interval, 0.083 to 0.674]; P=0.007) and OS (hazard ratio, 0.260 [95% confidence interval, 0.091 to 0.745] ; P=0.012). Conclusions: The prognostic impact of PD-L1 expression on tumor cells was distinct according to NLR. “NLR-low and tumoral PD-L1-negative” patients showed a favorable prognosis. Citation Format: Kazue Yoneda, Taiji Kuwata, Masataka Mori, Masatoshi Kanayama, Ayako Hirai, Yuko Tashima, Naoko Imanishi, Koji Kuroda, Yoshinobu Ichiki, Fumihiro Tanaka. Prognostic impact of PD-L1 expression in correlation with neutrophil-to-lymphocyte ratio in squamous cell carcinoma of the lung [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3149.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2019
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  • 7
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 74, No. 19_Supplement ( 2014-10-01), p. 3080-3080
    Abstract: Background. The CellSearch system is the most commonly used technique approved by the FDA to isolate and quantify circulation tumor cells (CTCs). However, in this system, the efficiency of CTC isolation is influenced by the epithelial cell adhesion molecule (EpCAM). Therefore, it is difficult to isolate CTCs in the setting of the epithelial to mesenchymal transition (EMT), malignant pleural mesothelioma and so on. We developed polymeric microfluidic devices for CTC isolation (CTC-chip) and compared the efficacy of CellSearch with CTC-chip using EpCAM-negative tumor cell lines of malignant pleural mesothelioma. Methods. In order to evaluate the accuracy of the number of CTCs, we used PBS samples and peripheral blood samples (blood samples) collected from healthy donors to which spiking the tumor cell lines ACC-MESO1 and ACC-MESO4. Both tumor cell lines (obtained from ATCC) were derived from malignant pleural mesothelioma, positive for mesothelin and podoplanin and negative for EpCAM. The recovery rate was evaluated using two methods. CellSearch was performed according to the normal method, while CTC-chip was performed using a two coating pattern with anti-mesothelin antibody (chip1) and anti-podoplanin antibody (chip2). Results. In the PBS samples, the recovery rate of captured ACC-MESO1 was 8.6%/98%/99% (CellSearch/chip 1/chip 2), while those of ACC-MESO4 was 21.1%/97%/99%, respectively. In the blood samples, the recovery rate of captured ACC-MESO1 was 1.2%/6.2%/13.3% (CellSearch/chip 1/chip 2), while that of ACC-MESO4 was 6.0%/3.7%/11.6%, respectively. Conclusions. Our experiments showed that the recovery rate of each sample was higher using CTC-chip than that obtained with CellSearch. Moreover, CTC-chip coated anti-podoplanin antibody exhibited a higher recovery rate than coated anti-mesothelin antibody. Future studies using experiments of CTC isolation in cancer patients with malignant pleural mesothelioma are thus required. Citation Format: Yasuhiro Chikaishi, Tomoko So, Masaru Takenaka, Soichi Oka, Ayako Hirai, Takashi Iwanami, Hidehiko Shimokawa, Kazue Yoneda, Yoshika Nagata, Hidetaka Uramoto, Takeshi Ohnaga, Fumihiro Tanaka. Comparison of CellSearch with polymeric microfluidic devices for CTC isolation using EpCAM-negative tumor cell lines of malignant pleural mesothelioma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3080. doi:10.1158/1538-7445.AM2014-3080
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2014
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    detail.hit.zdb_id: 1432-1
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  • 8
    Online Resource
    Online Resource
    American Association for Cancer Research (AACR) ; 2018
    In:  Cancer Research Vol. 78, No. 13_Supplement ( 2018-07-01), p. 1599-1599
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 78, No. 13_Supplement ( 2018-07-01), p. 1599-1599
    Abstract: Background: Circulating tumor cells (CTCs) are not only a surrogate of distant metastasis, but also useful for material of liquid biopsy. "Universal" CTC-chip is a polymeric microfluidic device that can capture CTCs according to their surface marker from whole blood. We have examined the capture efficiency of lung cancer and mesothelioma cells in the blood with CTC-chip coated with antibodies against various surface marker. Methods: Lung cancer cell lines (PC-9) or mesothelioma cell lines (ACC-MESO-4, MSTO-211H) were used in this study. Cells were CFSE-labeled and added in the blood of healthy donor. A polymeric CTC-chip was coated with two steps: base- and capture-antibody. As capture antibody, we used anti-EpCAM, podoplanin, and EGFR antibody. After flowing these samples, capture efficiency was calculated (number of captured cells/number of flowed cells). Results: Capture efficiency of PC-9, MESO-4, and METO-211H was 100.0%, 9.5% and 9.1% (EpCAM-chip; anti-EpCAM antibody clone HEA125); 5.8%, 85.9% and 9.0 % (podoplanin-chip; anti-podoplanin antibody clone E1); 38.6, 112.5%, 11.7% (podoplanin-chip; anti-podoplanin antibody clone NZ-1); 30.2%, 21.8%, 38.5% (EGFR-chip; anti-EGFR antibody clone 528), respectively. Conclusion: Using "Universal CTC-chip" with various antibodies, it was possible to capture nonepithelial tumor cells such as mesothelioma cells. Citation Format: Kazue Yoneda, Taiji Kuwata, Fumihiro Tanaka. Capture of tumor cells in blood with "Universal CTC-chip" [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1599.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
    RVK:
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2018
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    detail.hit.zdb_id: 1432-1
    detail.hit.zdb_id: 410466-3
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