Establishment of a monoclonal antibody PMab-233 for immunohistochemical analysis against Tasmanian devil podoplanin

Furusawa, Yoshikazu; Yamada, Shinji; Itai, Shunsuke; Nakamura, Takuro; Takei, Junko; Sano, M.; Harada, Hiroyuki; Fukui, Masato; Kaneko, Mika K.; Kato, Yukinari

doi:10.1016/j.bbrep.2019.100631

Cited by 64 publications

(37 citation statements)

References 42 publications

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“…One formalin-fixed paraffin-embedded (FFPE) tissue sample from an oropharyngeal squamous cell carcinoma patient who underwent surgery at Sendai Medical Center was used for this study ( 13 ). Written informed consent was obtained from the patient for sample procurement and subsequent data analyses.…”

Section: Methodsmentioning

confidence: 99%

“…CBIS has previously been used for developing several mAbs against various proteins, including the five transmembrane protein CD133 (7). Using the CBIS method, we have successfully produced sensitive and specific mAbs useful for not only flow cytometry, but also western blot and immunohistochemical analyses (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

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Establishment of C20Mab‑11, a novel anti‑CD20 monoclonal antibody, for the detection of B cells

2020

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CD20 is one of several B-lymphocyte antigens that has been shown to be an effective target for the detection and treatment of B-cell lymphomas. Sensitive and specific monoclonal antibodies (mAbs) are required for every application used for the diagnosis of B-cell lymphoma. Although many anti-CD20 mAbs have been established, the types of applications, those anti-CD20 can be used in, are limited. In this study, we aimed to establish novel anti-CD20 mAbs to be used for broad applications, such as flow cytometry, western blot, and immunohistochemical analyses, using the Cell-Based Immunization and Screening (CBIS) method. One of the established mAbs, C 20 Mab-11 (IgM, kappa), detected overexpression of CD20 in CHO-K1 or LN229 cell lines, indicating that C 20 Mab-11 is specific for CD20. In western blot analyses, C 20 Mab-11 detected not only overexpression of CD20 in CHO-K1 or LN229 cell lines, but also CD20 of BALL-1 and Raji cells with both sensitivity and specificity. Furthermore, C 20 Mab-11 strongly stained B cells of the lymph follicle and B cell lymphomas in immunohistochemical analyses. These results indicate that C 20 Mab-11 develped by CBIS method, is useful for the detection of CD20 in lymphoma tissues by flow cytometry, western blot, and immunohistochemical analyses and potentially could be beneficial for the treatment of B cell lymphomas.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Establishment of C20Mab‑11, a novel anti‑CD20 monoclonal antibody, for the detection of B cells

2020

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“…We have demonstrated that the critical epitope of PMab-237 could include Leu82 and Thr84 of wPDPN using the deletion mutants and point mutants of wPDPN in CHO-K1 cells by flow cytometry. In our previous studies, we developed many mAbs against PDPNs of human, (24) mouse, (25) rat, (26) rabbit, (27) dog, (28) cat, (29) bovine, (30) horse, (31) Tasmanian devil, (32) alpaca, (33) bear, (34) tiger, (35) goat, (36) pig, (37,38) and whale. (20) We successfully determined the binding epitope of those mAbs.…”

Section: Discussionmentioning

confidence: 99%

Epitope Analysis of an Anti-Whale Podoplanin Monoclonal Antibody, PMab-237, Using Flow Cytometry

Sayama

Sano

Kaneko

et al. 2020

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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Podoplanin (PDPN) is a small mucin-type transmembrane glycoprotein, which was first discovered in podocytes of the kidney. PDPN is a specific lymphatic endothelial marker and is also known as T1alpha, a marker of lung type I alveolar cells, or Aggrus, a platelet aggregation-inducing factor. PDPN possesses three platelet aggregationstimulating (PLAG) domains and PLAG-like domains (PLDs), which bind to C-type lectin-like receptor-2. Previously, we developed a novel anti-whale PDPN (wPDPN) monoclonal antibody (mAb) PMab-237 using the Cell-Based Immunization and Screening (CBIS) method and the RIEDL tag of Arg-Ile-Glu-Asp-Leu sequence. PMab-237 detected wPDPN by flow cytometry, western blot, and immunohistochemical analyses. However, the specific binding epitope of PMab-237 for wPDPN remains unknown. In this study, deletion mutants and point mutants of wPDPN with N-terminal RIEDL tag were produced to analyze the PMab-237 epitope using flow cytometry. The analysis of deletion mutants showed that the N-terminus of the PMab-237 epitope exists between the 80th amino acid (AA) and the 85th AA of wPDPN. In addition, the analysis of point mutants demonstrated that the critical epitope of PMab-237 includes Leu82 and Thr84 of wPDPN, indicating that the PMab-237 epitope is located in the PLD of wPDPN.

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“…The P3U1, CHO-K1, CHO/bPDPN, CHO/hPDPN [ 22 ], CHO/mouse PDPN (mPDPN) [ 22 ], CHO/rat PDPN (rPDPN) [ 10 ], CHO/rabbit PDPN (rabPDPN) [ 11 ], CHO/dog PDPN (dPDPN) [ 13 ], CHO/bovine PDPN (bovPDPN) [ 12 ], CHO/cat PDPN (cPDPN) [ 14 ], CHO/pig PDPN (pPDPN) [ 15 ], CHO/horse PDPN (horPDPN) [ 16 ], CHO/tiger PDPN (tigPDPN) [ 18 ], CHO/alpaca PDPN (aPDPN) [ 19 ], CHO/Tasmanian devil PDPN (tasPDPN) [ 21 ], CHO/goat PDPN (gPDPN) [ 17 ], CHO/sheep PDPN (sPDPN) [ 23 ], and CHO/whale PDPN (wPDPN) [ 20 ] were cultured in a Roswell Park Memorial Institute (RPMI) 1640 medium (Nacalai Tesque, Inc.), supplemented with 10% of heat-inactivated fetal bovine serum (FBS; Thermo Fisher Scientific Inc.), 100 units/mL of penicillin, 100 μg/mL of streptomycin, and 25 μg/mL of amphotericin B (Nacalai Tesque, Inc.). The cells were grown in an incubator at 37 °C with humidity and 5% CO 2 and 95% air atmosphere.…”

Section: Methodsmentioning

confidence: 99%

“…We have developed monoclonal antibodies (mAbs) against human [ 6 ], mouse [ 9 ], rat [ 10 ], rabbit [ 11 ], bovine [ 12 ], dog [ 13 ], cat [ 14 ], pig [ 15 ], horse [ 16 ], goat [ 17 ], tiger [ 18 ], alpaca [ 19 ], whale [ 20 ], and Tasmanian devil [ 21 ] PDPNs. In this study, we immunized mice with CHO/bear PDPN (bPDPN) cells and established mAbs against bPDPN.…”

Section: Introductionmentioning

confidence: 99%

Development of an anti-bear podoplanin monoclonal antibody PMab-247 for immunohistochemical analysis

Furusawa

Takei

Sayama

et al. 2019

Biochemistry and Biophysics Reports

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Sensitive and specific monoclonal antibodies (mAbs) targeting podoplanin (PDPN) are needed for immunohistochemical analyses using formalin-fixed paraffin-embedded tissues because PDPN is known as a lymphatic endothelial cell maker in pathology. Recently, we established anti -PDPN mAbs against many species, such as human, mouse, rat, rabbit, dog, cat, bovine, pig, horse, goat, tiger, alpaca, and Tasmanian devil. However, anti-bear PDPN (bPDPN) has not been established yet. In this study, we immunized mice with bPDPN-overexpressing Chinese hamster ovary (CHO)-K1 (CHO/bPDPN) cells, and screened mAbs against bPDPN using flow cytometry. One of the mAbs, PMab-247 (IgG 1 , kappa), specifically detected CHO/bPDPN cells by flow cytometry and immunohistochemistry. Our findings suggest the potential usefulness of PMab-247 for the functional analyses of bPDPN.

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Establishment of a monoclonal antibody PMab-233 for immunohistochemical analysis against Tasmanian devil podoplanin

Cited by 64 publications

References 42 publications

Establishment of C20Mab‑11, a novel anti‑CD20 monoclonal antibody, for the detection of B cells

Establishment of C20Mab‑11, a novel anti‑CD20 monoclonal antibody, for the detection of B cells

Epitope Analysis of an Anti-Whale Podoplanin Monoclonal Antibody, PMab-237, Using Flow Cytometry

Development of an anti-bear podoplanin monoclonal antibody PMab-247 for immunohistochemical analysis

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