Molecular cloning and chimerisation of CDI 315B monoclonal antibody.
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 VHand VLregions were joint to human IgG1and 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.
Department of Biochemistry and Molecular Biology, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia, , , , , , SI.
Nataša
N
Kopitar-Jerala
International Centre for Genetic Engineering and Biotechnology, Area Science Park, Padriciano 99, 34012 Trieste, Italy, , , , , , IT.
Marco
M
Bestagno
Department of Biochemistry and Molecular Biology, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia, , , , , , SI.
Xiaohui
X
Fan
Rudjer Bošković Institute, Bijenička cesta 54, 10000 Zagreb, Croatia, , , , , , HR.
Djurdja
D
Novak-Despot
International Centre for Genetic Engineering and Biotechnology, Area Science Park, Padriciano 99, 34012 Trieste, Italy, , , , , , IT.
Oscar
O
Burrone
Department of Biochemistry and Molecular Biology, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia, , , , , , SI.
Janko
J
Kos
Institute of Oncology, Zaloška cesta 2, 1105 Ljubljana, Slovenia, , , , , , SI.
Janez
J
Škrk
Department of Biochemistry and Molecular Biology, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia, , , , , , SI.
Franc
F
Gubenšek
Key words breast cancer; chimerisation.; recombinant antibodies