Establishment of Mouse Monoclonal Antibody LpMab-13 Against Human Podoplanin

OgasawaraSatoshi,; Kaneko, Mika K.; HonmaRyusuke,; OkiHiroharu,; FujiiYuki,; TakagiMichiaki,; SuzukiHiroyoshi,; Kato, Yukinari

doi:10.1089/mab.2016.0006

Cited by 17 publications

(8 citation statements)

References 42 publications

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“…This technology was also useful for generating anti-glycopeptide mAbs (GpMabs), including LpMab-3, (20) LpMab-9, (21) LpMab-12, (22) LpMab-19, (23) and LpMab-21. (24,25) Furthermore, we used this technology to generate mAbs that bind to various novel epitopes of podoplanin, including LpMab-7, (10)(11)(12) LpMab-10, (26) LpMab-13, (27) and LpMab-17. (28) We have also developed sensitive and specific anti-podocalyxin mAbs using the same methods.…”

Section: Discussionmentioning

confidence: 99%

H₂Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer

Itai

Fujii

Kaneko

et al. 2017

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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Human epidermal growth factor receptor 2 (HER2) plays a critical role in the progression of breast cancers, and HER2 overexpression is associated with poor clinical outcomes. Trastuzumab is an anti-HER2 humanized antibody that leads to significant survival benefits in patients with HER2-positive metastatic breast cancers. In this study, we developed novel anti-HER2 monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. Initially, we expressed the full length or ectodomain of HER2 in LN229 glioblastoma cells and then immunized mice with ectodomain of HER2 or LN229/HER2, and performed the first screening by enzyme-linked immunosorbent assays using ectodomain of HER2. Subsequently, we selected mAbs according to their efficacy in flow cytometry (second screening), Western blot (third screening), and immunohistochemical analyses (fourth screening). Among 100 mAb clones, only three mAbs reacted with HER2 in Western blot, and clone HMab-77 (IgG, kappa) was selected. Finally, immunohistochemical analyses with HMab-77 showed sensitive and specific reactions against breast cancer cells, warranting the use of HMab-77 to detect HER2 in pathological analyses of breast cancers.

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Section: Discussionmentioning

confidence: 99%

H₂Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer

Itai

Fujii

Kaneko

et al. 2017

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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“…LpMab-9, the epitope of which was identified as residues 25–30 of hPDPN [ 26 ], did not react with any glycopeptides of hpp3854. In contrast, LpMab-13 and LpMab-20, which were recently established using CasMab technology [ 30 ], recognized all the glycopepties of hpp3854. Collectively, these data indicate that the essential epitope of LpMab-12 is 49 -DVVT(SAα2-6GalNAc)P- 53 ( Fig 5 ).…”

Section: Resultsmentioning

confidence: 99%

LpMab-12 Established by CasMab Technology Specifically Detects Sialylated O-Glycan on Thr52 of Platelet Aggregation-Stimulating Domain of Human Podoplanin

et al. 2016

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Podoplanin (PDPN), also known as Aggrus, possesses three tandem repeat of platelet aggregation-stimulating (PLAG) domains in its N-terminus. Among the PLAG domains, sialylated O-glycan on Thr52 of PLAG3 is essential for the binding to C-type lectin-like receptor-2 (CLEC-2) and the platelet-aggregating activity of human PDPN (hPDPN). Although various anti-hPDPN monoclonal antibodies (mAbs) have been generated, no specific mAb has been reported to target the epitope containing glycosylated Thr52. We recently established CasMab technology to develop mAbs against glycosylated membrane proteins. Herein, we report the development of a novel anti-glycopeptide mAb (GpMab), LpMab-12. LpMab-12 detected endogenous hPDPN by flow cytometry. Immunohistochemical analyses also showed that hPDPN-expressing lymphatic endothelial and cancer cells were clearly labeled by LpMab-12. The minimal epitope of LpMab-12 was identified as Asp49–Pro53 of hPDPN. Furthermore, LpMab-12 reacted with the synthetic glycopeptide of hPDPN, corresponding to 38–54 amino acids (hpp3854: 38-EGGVAMPGAEDDVVTPG-54), which carries α2–6 sialylated N-acetyl-D-galactosamine (GalNAc) on Thr52. LpMab-12 did not recognize non-sialylated GalNAc-attached glycopeptide, indicating that sialylated GalNAc on Thr52 is necessary for the binding of LpMab-12 to hPDPN. Thus, LpMab-12 could serve as a new diagnostic tool for determining whether hPDPN possesses the sialylation on Thr52, a site-specific post-translational modification critical for the hPDPN association with CLEC-2.

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“…An anti-human podoplanin antibody (NZ-1) inhibiting podoplanin-induced platelet aggregation, abrogated experimental metastasis has already been formed, however with strong toxic side effects. A further novel chimeric humanized anti-human podoplanin antibody inhibiting podoplanin-induced platelet aggregation has been developed as a potential novel anticancer agent[ 39 ]. Since there is toxicity due to interferences with endogenous podoplanin in other cell types like type I lung alveolar cells, kidney podocytes, choroid plexus epithelium, a less interfering antigen has been developed[ 40 ].…”

Section: Discussionmentioning

confidence: 99%

Neoadjuvant chemoradiation changes podoplanin expression in esophageal cancer patients

Warnecke-Eberz¹,

Plum²,

Schweinsberg³

et al. 2020

WJG

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BACKGROUND Locally advanced adenocarcinoma of the esophagus (EAC) and squamous cell carcinoma (ESCC) result in a worse prognosis. Neoadjuvant treatment improves survival, however, only for responders. The transmembrane glycoprotein podoplanin is overexpressed in squamous cell carcinomas, miRNA-363 is associated to its regulation in head and neck cancer. AIM To predict therapy response and prognosis markers, and targets for novel therapies would individualize treatments leading to more favourable outcomes. METHODS Expression of podoplanin protein has been visualized by immunohistochemistry in surgical specimens of 195 esophageal cancer patients who underwent transthoracic esophagectomy: 90 ESCC and 105 EAC with clinical T2-3, Nx, M0. One hundred and six patients received neoadjuvant chemoradiation. RNA was extracted from paraffin-embedded tissue, and miRNA-363 quantified by real-time TaqMan-real-time-PCR. D2-40 mab staining of > 5% was scored as high podoplanin expression (HPE). We related podoplanin and miRNA-363 expression to histopathologic response after neoadjuvant treatment and clinicopathological characteristics, such as histological tumor type, survival rate or clinical tumor category. RESULTS We confirmed expression of membrane-bound podoplanin in 90 ESCC patients. 26% showed HPE of > 5%. In addition, absence in EAC patients (only 2% with HPE) was shown. Lower podoplanin expression has been detected in resection-specimen of 58 ESCC patients after neoadjuvant (RTx/CTx) treatment, only 11% with HPE, compared to 50% HPE of 32 non-pretreated primary surgery patients, P = 0.0001. This difference of podoplanin expression was confirmed comparing pre-treatment biopsies with matching post-treatment surgical specimens, P < 0.001. Podoplanin has been identified as a prognostic marker in 32 patients that underwent primary surgery without neoadjuvant treatment. Low (0-5%) podoplanin expression was associated with better prognosis compared to patients with HPE, P = 0.013. Podoplanin expression has been associated with post-transcriptional regulation by miRNA-363. At a cut-off value of miR-363 < 7, lower miR-363 expression correlated with HPE in surgical tissue specimens of primary surgery patients, P = 0.013. Therefore, ESCC patients with miRNA-363 expression < 7 had a worse prognosis than patients expressing miRNA-363 ≥ 7, P = 0.049. CONCLUSION Analysis of the molecular process that leads to decrease in podoplanin expression during neoadjuvant treatment and its regulation may provide novel markers and targets to improve targeted therapy of ESCC.

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Establishment of Mouse Monoclonal Antibody LpMab-13 Against Human Podoplanin

Cited by 17 publications

References 42 publications

H₂Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer

H₂Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer

LpMab-12 Established by CasMab Technology Specifically Detects Sialylated O-Glycan on Thr52 of Platelet Aggregation-Stimulating Domain of Human Podoplanin

Neoadjuvant chemoradiation changes podoplanin expression in esophageal cancer patients

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Establishment of Mouse Monoclonal Antibody LpMab-13 Against Human Podoplanin

Cited by 17 publications

References 42 publications

H2Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer

H2Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer

LpMab-12 Established by CasMab Technology Specifically Detects Sialylated O-Glycan on Thr52 of Platelet Aggregation-Stimulating Domain of Human Podoplanin

Neoadjuvant chemoradiation changes podoplanin expression in esophageal cancer patients

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H₂Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer

H₂Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer