Quantitative autoradiography to determine local GD2 antigen concentration of a human neuroblastoma xenograft growing in nude mice using monoclonal antibody 3F8

L. Kostakoglu, O. Ugur, N. K.V. Cheung, J. Zhang, Y. Huang, S. M. Larson

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Abstract

We studied the antigen density (B(max) and binding affinity (K(a)) of the monoclonal antibody (MAb) 3F8, a murine IgG3, selective for ganglioside GD2, to the neuroblastoma tumor in order to further understand the influence of the above factors on the antibody-antigen systems. We compared the results of B(max) and K(a) obtained from quantitative autoradiography (QAR) of xenografted NMB-7 tumors with competitive inhibition of in vitro binding assay (CIBA) using NMB-7 cell suspension, two situations with different cell growth characteristics. Both methods yielded a K(a) value of 1-2 x 108 L M-1, an antibody binding affinity which is not exceptional and shared by many murine anti-tumor antibodies. On the other hand, the mean maximal concentration, B(m)ax, of GD2 antigen in tumor tissue was 3423 ± 357 picomols g-1 by QAR. This value is approximately 10 fold higher than reported with other antigen systems such as p97 and the chondroitin sulfate proteoglycan. CIBA revealed a mean B(m)ax of 15 ± 2.8 x 106 sites of GD2 antigen per cell (12534 ± 2300 picomol g) assuming bivalent binding.

Original languageEnglish
Pages (from-to)145-154
Number of pages10
JournalAntibody, Immunoconjugates, and Radiopharmaceuticals
Volume8
Issue number3
StatePublished - 1995
Externally publishedYes

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