Generation and characterization of a mouse/human chimeric antibody directed against extracellular matrix protein tenascin

Xuanmin He, Gary E. Archer, Carol J. Wikstrand, Sherie L. Morrison, Michael R. Zalutsky, Darell D. Bigner, Surinder K. Batra

Research output: Contribution to journalArticle

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Abstract

The murine anti-tenascin monoclonal antibody 81C6, following iodination, has been shown to be an efficient localizing and therapeutic agent in both subcutaneous and intracranial human glioma xenograft models in athymic mice and rats. Similarly, effective monoclonal antibody 81C6 localization has been demonstrated in glioma patients, and Phase I trials with the intact murine IgG2b κ molecule are currently in progress. In order to maximize the potential for repeated administration by minimizing murine Fc-mediated immunogenicity and reducing Fc-mediated immune effects, we created murine 81C6 variable region/human IgG2 chimeric monoclonal antibodies by the molecular cloning of the variable region genes of mouse 81C6 and their genetic linkage to human constant region exons. The resulting chimeric constructs were introduced into SP 2 0 cells, and stable transfectomas were selected by G418 and mycophenolic acid resistance. The resistant clones were screened for anti-tenascin activity on tenascin-coated plates by enzyme-linked immunosorbent assay. The N-terminal amino acid sequence of both heavy and light chains of the purified chimeric 81C6 antibody matched exactly with that of the native mouse 81C6 as well as with that deduced from the nucleotide sequence. The production level of chimeric 81C6 (13.9 mg/ml) from ascites in the highest expressing transfectoma was much higher than that of native mouse 81C6 (2.5 mg/ml). The chimeric antibody showed the same specificity and equivalent affinity for human intact tenascin or tenascin-expressing cells as the native mouse 81C6 antibody. Direct comparison of radioiodinated chimeric and radioiodinated mouse 81C6 biodistribution in subcutaneous and intracranial xenograft-bearing mice showed higher tumor-to-normal tissue ratios for chimeric 81C6 as compared with native mouse 81C6. The improved localizing and clearance characteristics of chimeric 81C6 in xenograft model systems suggests that chimeric 81C6 would be an improved reagent for intracompartmental therapy of tenascin-expressing tumors in the human central nervous system.

Original languageEnglish (US)
Pages (from-to)127-137
Number of pages11
JournalJournal of Neuroimmunology
Volume52
Issue number2
DOIs
StatePublished - Jul 1994

Fingerprint

Tenascin
Extracellular Matrix Proteins
Antibodies
Heterografts
Monoclonal Antibodies
Glioma
Nude Rats
Mycophenolic Acid
Genetic Linkage
Halogenation
Molecular Cloning
Ascites
Nude Mice
Amino Acid Sequence
Exons
Neoplasms
Central Nervous System
Clone Cells
Immunoglobulin G
Enzyme-Linked Immunosorbent Assay

Keywords

  • Chimeric antibody
  • Extracellular matrix protein
  • Molecular cloning
  • Neoplasm

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Neurology
  • Clinical Neurology

Cite this

Generation and characterization of a mouse/human chimeric antibody directed against extracellular matrix protein tenascin. / He, Xuanmin; Archer, Gary E.; Wikstrand, Carol J.; Morrison, Sherie L.; Zalutsky, Michael R.; Bigner, Darell D.; Batra, Surinder K.

In: Journal of Neuroimmunology, Vol. 52, No. 2, 07.1994, p. 127-137.

Research output: Contribution to journalArticle

He, Xuanmin ; Archer, Gary E. ; Wikstrand, Carol J. ; Morrison, Sherie L. ; Zalutsky, Michael R. ; Bigner, Darell D. ; Batra, Surinder K. / Generation and characterization of a mouse/human chimeric antibody directed against extracellular matrix protein tenascin. In: Journal of Neuroimmunology. 1994 ; Vol. 52, No. 2. pp. 127-137.
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