TY - JOUR
T1 - Establishment of monoclonal anti-human CD26 antibodies suitable for immunostaining of formalin-fixed tissue
AU - Hatano, Ryo
AU - Yamada, Taketo
AU - Matsuoka, Shuji
AU - Iwata, Satoshi
AU - Yamazaki, Hiroto
AU - Komiya, Eriko
AU - Okamoto, Toshihiro
AU - Dang, Nam H.
AU - Ohnuma, Kei
AU - Morimoto, Chikao
N1 - Funding Information:
The authors thank Ms. Hiroko Madokoro for excellent assistance with immunohistochemical staining of CD26, and also thank Ms. Haruna Otsuka and Ms. Aya Miwa for the assistance with preparation of soluble CD26 and the experiment of epitope mapping. This work was supported by Grant-in-Aid of The Ministry of Education, Science, Sports (KO and CM) and Culture, Ministry of Health, Labour, and Welfare, Japan (CM).
PY - 2014/1
Y1 - 2014/1
N2 - Background: A T cell costimulatory molecule with dipeptidyl peptidase IV (DPPIV) activity in its extracellular region, CD26 is a multifunctional molecule associated with various proteins such as adenosine deaminase, caveolin-1, CXCR4, collagen, and fibronectin, while playing an important role in the regulation of inflammatory responses and tumor biology. We have focused on CD26 as a novel therapeutic target for various tumors and immune disorders, and have developed a humanized anti-CD26 monoclonal antibody (mAb), YS110, which is currently being evaluated in a phase I clinical trial for patients with CD26-expressing tumors, including malignant mesothelioma. Since detection of tumor CD26 expression is required for determining potential eligibility for YS110 therapy, the development of anti-human CD26 mAb that can clearly and reliably detect the denatured CD26 molecule in the formalin-fixed paraffin-embedded tissues is critical.Methods: To develop novel anti-CD26 mAbs capable of binding to the denatured CD26, we immunized mice with CD26 protein denatured in urea buffer. After the fusion of splenocytes and myeloma cells, the mAbs were screened for specific reactivity with human CD26 by flow cytometry, enzyme-linked immunosorbent assay, and immunohistochemistry. The binding competitiveness of novel anti-CD26 mAbs with the humanized anti-CD26 mAb YS110 was also examined.Results: We have succeeded in developing novel anti-human CD26 mAbs suitable for immunohistochemical staining of CD26 in formalin-fixed tissue sections with reliable clarity and intensity. Importantly, some of these mAbs exhibit no cross-reactivity with the humanized anti-CD26 mAb.Conclusions: These novel mAbs are potentially useful as companion diagnostic agents to analyze CD26 expression in the clinical setting while advancing future CD26-related research.
AB - Background: A T cell costimulatory molecule with dipeptidyl peptidase IV (DPPIV) activity in its extracellular region, CD26 is a multifunctional molecule associated with various proteins such as adenosine deaminase, caveolin-1, CXCR4, collagen, and fibronectin, while playing an important role in the regulation of inflammatory responses and tumor biology. We have focused on CD26 as a novel therapeutic target for various tumors and immune disorders, and have developed a humanized anti-CD26 monoclonal antibody (mAb), YS110, which is currently being evaluated in a phase I clinical trial for patients with CD26-expressing tumors, including malignant mesothelioma. Since detection of tumor CD26 expression is required for determining potential eligibility for YS110 therapy, the development of anti-human CD26 mAb that can clearly and reliably detect the denatured CD26 molecule in the formalin-fixed paraffin-embedded tissues is critical.Methods: To develop novel anti-CD26 mAbs capable of binding to the denatured CD26, we immunized mice with CD26 protein denatured in urea buffer. After the fusion of splenocytes and myeloma cells, the mAbs were screened for specific reactivity with human CD26 by flow cytometry, enzyme-linked immunosorbent assay, and immunohistochemistry. The binding competitiveness of novel anti-CD26 mAbs with the humanized anti-CD26 mAb YS110 was also examined.Results: We have succeeded in developing novel anti-human CD26 mAbs suitable for immunohistochemical staining of CD26 in formalin-fixed tissue sections with reliable clarity and intensity. Importantly, some of these mAbs exhibit no cross-reactivity with the humanized anti-CD26 mAb.Conclusions: These novel mAbs are potentially useful as companion diagnostic agents to analyze CD26 expression in the clinical setting while advancing future CD26-related research.
KW - CD26/dipeptidyl peptidase 4
KW - Companion diagnostic drug
KW - Immunohistochemical staining
KW - Malignant mesothelioma
KW - T cell costimulation
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U2 - 10.1186/1746-1596-9-30
DO - 10.1186/1746-1596-9-30
M3 - Article
C2 - 24502396
AN - SCOPUS:84897711744
SN - 1746-1596
VL - 9
JO - Diagnostic Pathology
JF - Diagnostic Pathology
IS - 1
M1 - 30
ER -