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Title: The production and preclinical characterization of a chimeric anti-breast-cancer antibody, cBC2.
Austin Authors: Sutton, V R;Burgess, John R;Pietersz, Geoffrey A;Li, W J;McKenzie, Ian F C;Trapani, Joseph A
Affiliation: Austin Research Institute, Austin Hospital, Heidelberg, Victoria, Australia
Issue Date: 1-Apr-1994
Publication information: Therapeutic Immunology; 1(2): 83-93
Abstract: A chimeric (mouse-human) BC2 antibody (cBC2) was produced which may be used in the diagnosis and treatment of breast cancer. The BC2 variable region genes were amplified by polymerase chain reaction (PCR), using oligonucleotide primers homologous to the framework sequences of mouse VH and V kappa genes. The PCR products were used to create cBC2 expression vectors containing the mouse BC2 VH and V kappa and human constant region (IgG1 and K) genes. Chimeric antibody was produced following transfection of these constructs into Sp2/0 myeloma cells. Binding assays in vitro demonstrated that cBC2 had the same specificity for human milk fat globule membrane (HMFGM) and MUC1+ cells as mBC2, and bound antigen with a similar affinity (cBC2, Ka 5.53 +/- 2.09 x 10(8); mBC2, Ka 1.44 +/- 0.98 x 10(9)). Functionally, only cBC2 (5-25 micrograms ml-1), was able to mediate antibody-dependent cellular cytotoxicity (ADCC) with human effector cells, with 25% maximal specific lysis of MUC1+ cells at an E/T ratio of 100:1. Human complement-mediated lysis was minimal (10-15% specific lysis) with both mBC2 and cBC2. Neither cBC2 nor mBC2 was able to inhibit tumour growth in vivo in the absence of covalently coupled anticancer drugs. However, biodistribution studies demonstrated that both antibodies preferentially targeted MUC1+ tumour cells, with 17% of the injected dose of cBC2, as compared to 27% of mBC2, localized to the MUC1+ tumour at 24 h (less than 6% detected in any other tissue).
Gov't Doc #: 7584487
Type: Journal Article
Subjects: 3T3 Cells
Amino Acid Sequence
Antibodies, Monoclonal.genetics
Antibodies, Neoplasm.genetics.immunology
Antibody Affinity
Antibody-Dependent Cell Cytotoxicity
Antigens, Neoplasm.immunology
Base Sequence
Breast Neoplasms.immunology
Cloning, Molecular
Genes, Immunoglobulin
Immunoglobulin Heavy Chains.genetics
Immunoglobulin kappa-Chains.genetics
Mammary Neoplasms, Experimental.immunology
Mice, Inbred BALB C
Molecular Sequence Data
Recombinant Fusion Proteins
Tissue Distribution
Appears in Collections:Journal articles

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