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Monoclonal antibody 4C4‐mAb specifically recognizes keratan sulphate proteoglycan on human embryonal carcinoma cells
Author(s) -
Fukuma Mariko,
Abe Hitoshi,
Okita Hajime,
Yamada Taketo,
Hata Junichi
Publication year - 2003
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.1403
Subject(s) - monoclonal antibody , antigen , embryonal carcinoma , microbiology and biotechnology , proteoglycan , keratan sulfate , teratocarcinoma , biology , somatic cell , antibody , chemistry , cellular differentiation , biochemistry , immunology , extracellular matrix , gene
Germ cell tumours, the most common solid cancers in young males, display pluripotentiality for embryonal and somatic differentiation. Specific surface antigens are useful in the study of cellular differentiation and for clinical diagnosis. A mouse monoclonal antibody (4C4‐mAb) has been developed against a human embryonal carcinoma (EC) cell line (NCR‐G3) isolated from a combined form of testicular germ cell tumour. On immunohistological and immuno‐electron microscopic examination, the 4C4 antigen (4C4) was detected on the surface of NCR‐G3 and gold particles were exclusively detected on the microvilli of the cells. In both formalin‐fixed paraffin wax sections and touch‐smear specimens, 4C4 was detected specifically in EC, while the antigen was not expressed in other types of germ cell tumour or in the other solid tumours tested. Tunicamycin diminished the antigenicity of NCR‐G3 cells. In biochemical studies, 4C4 was found in a high molecular weight region ranging from 1 × 10 6 to 1 × 10 7 kD, which disappeared after periodate treatment. The density of 4C4 was 1.5 g/cm 3 after equilibrium centrifugation. These results imply that 4C4 is a proteoglycan. Furthermore, endo‐ and exo‐glycosidase treatment revealed that 4C4 is a keratan sulphate proteoglycan that contains sialyl and fucosyl moieties. With EC‐specific and formalin‐resistant characteristics, 4C4 may be a specific marker for diagnosing EC among a variety of germ cell tumours. Copyright © 2003 John Wiley & Sons, Ltd.