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Molecular cloning and chimerisation of CDI 315B monoclonal antibody.

Authors
  • Kopitar-Jerala, Nataša1
  • Bestagno, Marco2
  • Fan, Xiaohui1
  • Novak-Despot, Djurdja3
  • Burrone, Oscar2
  • Kos, Janko1
  • Škrk, Janez4
  • Gubenšek, Franc1
  • 1 Department of Biochemistry and Molecular Biology, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia, , , , , , SI. , (Slovenia)
  • 2 International Centre for Genetic Engineering and Biotechnology, Area Science Park, Padriciano 99, 34012 Trieste, Italy, , , , , , IT. , (Italy)
  • 3 Rudjer Bošković Institute, Bijenička cesta 54, 10000 Zagreb, Croatia, , , , , , HR. , (Croatia)
  • 4 Institute of Oncology, Zaloška cesta 2, 1105 Ljubljana, Slovenia, , , , , , SI. , (Slovenia)
Type
Published Article
Journal
Pflügers Archiv - European Journal of Physiology
Publisher
Springer-Verlag
Publication Date
Jan 01, 2000
Volume
439
Issue
Suppl 1
Identifiers
DOI: 10.1007/s004240000098
PMID: 28176080
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

A chimeric mouse-human antibody has been created that recognizes an antigen found on breast cancer cells and melanoma cells. Immunoglobulin constant domains of mouse monoclonal antibody CDI 315B Cγ1 and Cκ, were substituted by the human Cγ1 and Cκ. The CDI 315B variable heavy and light chain regions were PCR amplified from hybridoma RNA and sequenced. Mouse variable VH and VL regions were joint to human IgG1 and κ constant regions and subcloned into pcDNA3 expression vectors. The Sp2/0 murine myeloma cells were transfected with expression vectors pcDNA3L and pcDNA3H and the reactivity of chimeric antibodies was tested by indirect ELISA using B16F1 murine melanoma cells as well as MCF7 human breast cancer cells, as antigen.

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