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Anti-neuroblastoma antibody chCE7 binds to an isoform of L1-CAM present in renal carcinoma cells

✍ Scribed by Marina L. Meli; François Carrel; Robert Waibel; Hanspeter Amstutz; Nigel Crompton; Rolf Jaussi; Holger Moch; P. August Schubiger; Ilse Novak-Hofer


Publisher
John Wiley and Sons
Year
1999
Tongue
French
Weight
231 KB
Volume
83
Category
Article
ISSN
0020-7136

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✦ Synopsis


Immunoprecipitation after cell surface labeling of human neuroblastoma cells showed that the anti-neuroblastoma monoclonal antibody (mAb) chCE7 binds to a 200,000 M r cell surface protein. The protein was partially purified by immunoaffinity chromatography from a human renal carcinoma and a human neuroblastoma cell line, which both showed high levels of binding of MAb chCE7. NH 2 -terminal sequences of 18 and 15 amino acid residues were determined. Both sequences isolated from the renal carcinoma and the neuroblastoma cells showed strong homology to human cell adhesion molecule L1 (L1-CAM), and both were characterized by the NH 2 -terminal deletion of 5 amino acids, comprising exon 2 of L1-CAM. Reverse trancription-polymerase chain reaction (RT-PCR) analysis of the regions spanning exon 2 and exon 27 of L1-CAM indicated that in neuroblastoma cells both transcripts for the full-length and exon-deleted forms are present, whereas in the renal carcinoma cell lines only the exon-deleted L1-CAM isoform were detected. Western blot analysis showed that 6 of 7 tested renal carcinoma cell lines and 5 of 15 renal carcinoma tissues expressed L1-CAM. In normal adult kidney tissue, very low levels of protein expression were found. Northern blot analysis confirmed that in renal carcinoma and neuroblastoma cell lines L1-CAM mRNA levels are correlated with protein expression. Int.