Generation of monoclonal antibodies to cancer/testis (CT) antigen CT10/MAGE-C2.
CT10/MAGE-C2 is a recently identified antigen that, typically of cancer/testis (CT) antigens, can be found in various malignant tumors and in normal adult testis. As with many other CT antigens, our knowledge is based mainly on mRNA expression data. In the present study, we describe the generation of mAbs to CT10/MAGE-C2 for the analysis of its protein expression. Newly generated clones were chosen based on their reactivity in ELISA, immunoblotting, and immunohistochemistry (IHC). Emphasis was put on the reactivity of newly generated reagents on formalin-fixed, paraffin-embedded tissue to ensure their applicability to archival material. Eventually we selected two clones, LX-CT10.5 and LX-CT10.9, that showed intense reactivity to CT10/MAGE-C2 protein and CT10/MAGE-C2 mRNA-positive cell lines, but no cross-reactivity with other CT antigens. Both mAbs show superior staining characteristics in IHC and are applicable to frozen and paraffin sections. In testis, CT10/MAGE-C2 displays the typical CT pattern with regard to staining of germ cells, which is intense during the early maturation stages. In tumors, we analyzed a limited number of cases displaying the typical heterogeneous CT expression pattern. Interestingly, immunoreactivity was seen solely in the nucleus: No staining was seen in the cytoplasm of tumor cells.
Zhuang
Ran
R
Department of Immunology, Fourth Military Medical University, Xi'an 710032, PR China.
Zhu
Yong
Y
Fang
Liang
L
Liu
Xue-Song
XS
Tian
Ying
Y
Chen
Li-Hua
LH
Ouyang
Wei-Ming
WM
Xu
Xiao-Guang
XG
Jian
Jin-Long
JL
Güre
Ali O
AO
Fortunato
Sheila
S
Ritter
Gerd
G
Old
Lloyd J
LJ
Simpson
Andrew J G
AJ
Chen
Yao T
YT
Jin
Boquan
B
Jungbluth
Achim A
AA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000951:Antigens, Neoplasm; D002467:Cell Nucleus; D003593:Cytoplasm; D004797:Enzyme-Linked Immunosorbent Assay; D020869:Gene Expression Profiling; D006801:Humans; D006825:Hybridomas; D007150:Immunohistochemistry; D008297:Male; D008545:Melanoma / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000276:immunology*; D012878:Skin Neoplasms / Q000473:pathology; D013154:Spleen / Q000166:cytology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; C405429:MAGEC2 protein, human; D009363:Neoplasm Proteins