A Defucosylated Anti-EpCAM Monoclonal Antibody (EpMab-37-mG2a-f) Exerts Antitumor Activity in Xenograft Model

Asano, Teizo; Tanaka, Tomohiro; Suzuki, Hiroyuki; Li, Guanjie; Ohishi, Tomokazu; Kawada, Manabu; Yoshikawa, Takeo; Kaneko, Mika K.; Kato, Yukinari

doi:10.3390/antib11040074

Cited by 15 publications

(15 citation statements)

References 63 publications

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“…To evaluate the in vivo effect, we previously converted the IgG 1 subclass of mAbs into a mouse IgG 2a , and produced a defucosylated version. These defucosylated IgG 2a mAbs exhibited potent ADCC in vitro, and reduced tumor growth in mouse xenograft models [ 24 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ]. Therefore, the production of a class-switched and defucosylated version of C 44 Mab-9 is required to evaluate the antitumor activity in vivo.…”

Section: Discussionmentioning

confidence: 99%

Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C44Mab-9 for Multiple Applications against Colorectal Carcinomas

Ejima

Suzuki

Tanaka

et al. 2023

IJMS

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CD44 is a cell surface glycoprotein, and its isoforms are produced by the alternative splicing with the standard and variant exons. The CD44 variant exon-containing isoforms (CD44v) are overexpressed in carcinomas. CD44v6 is one of the CD44v, and its overexpression predicts poor prognosis in colorectal cancer (CRC) patients. CD44v6 plays critical roles in CRC adhesion, proliferation, stemness, invasiveness, and chemoresistance. Therefore, CD44v6 is a promising target for cancer diagnosis and therapy for CRC. In this study, we established anti-CD44 monoclonal antibodies (mAbs) by immunizing mice with CD44v3-10-overexpressed Chinese hamster ovary (CHO)-K1 cells. We then characterized them using enzyme-linked immunosorbent assay, flow cytometry, western blotting, and immunohistochemistry. One of the established clones (C44Mab-9; IgG1, kappa) reacted with a peptide of the variant 6-encoded region, indicating that C44Mab-9 recognizes CD44v6. Furthermore, C44Mab-9 reacted with CHO/CD44v3-10 cells or CRC cell lines (COLO201 and COLO205) by flow cytometry. The apparent dissociation constant (KD) of C44Mab-9 for CHO/CD44v3-10, COLO201, and COLO205 was 8.1 × 10−9 M, 1.7 × 10−8 M, and 2.3 × 10−8 M, respectively. C44Mab-9 detected the CD44v3-10 in western blotting, and partially stained the formalin-fixed paraffin-embedded CRC tissues in immunohistochemistry. Collectively, C44Mab-9 is useful for detecting CD44v6 in various applications.

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Section: Discussionmentioning

confidence: 99%

Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C44Mab-9 for Multiple Applications against Colorectal Carcinomas

Ejima

Suzuki

Tanaka

et al. 2023

IJMS

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“…Our group converted mouse IgG1 subclass of mAbs into a IgG2a, and produced a defucosylated mAbs using fucosyltransferases 8-deficient CHO-K1 cells. The defucosylated IgG2a mAbs showed potent antibody-dependent cellular cytotoxicity in vitro, and suppressed the tumor xenograft growth [27,[59][60][61][62][63][64][65]. Therefore, the production of a classswitched and defucosylated version of C44Mab-3 is required to evaluate the antitumor activity in vivo.…”

Section: Discussionmentioning

confidence: 99%

Development of a Novel Anti-CD44 Variant 5 Monoclonal Antibody C<sub>44</sub>Mab-3 for Multiple Applications against Pancreatic Carcinomas

Kudo¹,

Suzuki²,

Tanaka³

et al. 2023

Preprint

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Pancreatic cancer exhibits a poor prognosis due to the lack of early diagnostic biomarkers and the resistance to conventional chemotherapy. CD44 has been known as a cancer stem cell marker, and plays tumor promotion and drug resistance in various cancers. Especially, the splicing variants are overexpressed in many carcinomas, and play essential roles in the cancer stemness, invasiveness or metastasis, and resistance to treatments. Therefore, the understanding of each CD44 variant (CD44v) function and distribution in carcinomas is essential for the establishment of CD44-targeting tumor therapy. In this study, we immunized mice with CD44v3–10-overexpressed Chinese hamster ovary-K1 (CHO) cells, and established various anti-CD44 monoclonal antibodies (mAbs). One of the established clones (C44Mab-3; IgG1, kappa) recognized peptides of the variant 5-encoded region, indicating that C44Mab-3 is a specific mAb for CD44v5. Moreover, C44Mab-3 reacted with CHO/CD44v3–10 cells or pancreatic cancer cell lines (PK-1 and PK-8) by flow cytometry. The apparent KD of C44Mab-3 for CHO/CD44v3–10 and PK-1 was 7.1 × 10−10 M and 1.9 × 10−9 M, respectively. C44Mab-3 could detect the exogenous CD44v3–10 and endogenous CD44v5 in western blotting, and stained the formalin-fixed paraffin-embedded pancreatic cancer cells, but not normal pancreatic epithelial cells in immunohistochemistry. These results indicate that C44Mab-3 is useful for detecting CD44v5 in various applications, and expected for the application of pancreatic cancer diagnosis and therapy.

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“…To evaluate the in vivo effect, we previously converted the IgG1 subclass of mAbs into a mouse IgG2a, and produced a defucosylated version. These defucosylated IgG2a mAbs exhibited potent antibody dependent cellular cytotoxicity in vitro, and reduced the tumor growth in mouse xenograft models [38][39][40][41][42][43][44]. Therefore, the production of a class switched and defucosylated version of C44Mab-9 is required to evaluate the antitumor activity in vivo.…”

Section: Discussionmentioning

confidence: 99%

Development of a Novel Anti-CD44 variant 6 Monoclonal Antibody for Multiple Applications against Colorectal Carcinomas

Ejima¹,

Suzuki²,

Tanaka³

et al. 2023

Preprint

View full text Add to dashboard Cite

CD44 is a cell surface glycoprotein, and its isoforms are produced by the alternative splicing with the standard and variant exons. The CD44 variant exon containing isoforms (CD44v) are overexpressed in carcinomas. CD44v6 is one of the CD44v, and its overexpression predicts poor prognosis in colorectal cancer (CRC) patients. CD44v6 plays critical roles in CRC adhesion, proliferation, stemness, invasiveness, and chemoresistance. Therefore, CD44v6 is a promising target for cancer diagnosis and therapy for CRC. In this study, we established anti-CD44 monoclonal antibodies (mAbs) by immunizing mice with CD44v3-10-overexpressed Chinese hamster ovary-K1 (CHO) cells. We then characterized them using enzyme-linked immunosorbent assay, flow cytometry, western blotting, and immunohistochemistry. One of the established clones (C44Mab-9; IgG1, kappa) reacted with a peptide of variant 6-encoded region, indicating that C44Mab-9 recognizes CD44v6. Furthermore, C44Mab-9 reacted with CHO/CD44v3-10 cells or CRC cell lines (COLO201 and COLO205) by flow cytometry. The apparent KD of C44Mab-9 for CHO/CD44v3-10, COLO201, and COLO205 was 8.1 × 10−9 M, 1.7 × 10−8 M, and 2.3 × 10−8 M, respectively. C44Mab-9 detected the CD44v3-10 in western blotting, and partially stained the formalin-fixed paraffin-embedded CRC tissues in immunohistochemistry. Collectively, C44Mab-9 is useful for detecting CD44v6 in various applications.

show abstract

A Defucosylated Anti-EpCAM Monoclonal Antibody (EpMab-37-mG2a-f) Exerts Antitumor Activity in Xenograft Model

Cited by 15 publications

References 63 publications

Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C44Mab-9 for Multiple Applications against Colorectal Carcinomas

Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C44Mab-9 for Multiple Applications against Colorectal Carcinomas

Development of a Novel Anti-CD44 Variant 5 Monoclonal Antibody C<sub>44</sub>Mab-3 for Multiple Applications against Pancreatic Carcinomas

Development of a Novel Anti-CD44 variant 6 Monoclonal Antibody for Multiple Applications against Colorectal Carcinomas

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