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The development and purification of a bispecific antibody for lymphokine-activated killer cell targeting against the rat colon carcinoma CC531.
- Source :
-
Cancer immunology, immunotherapy : CII [Cancer Immunol Immunother] 1993 Jun; Vol. 36 (6), pp. 403-8. - Publication Year :
- 1993
-
Abstract
- In vivo targeting of lymphokine-activated killer (LAK) cells to tumour deposits by bispecific monoclonal antibodies (bimAb) may be a way to improve adoptive immunotherapy. We developed a bimAb against adherent LAK (ALAK) cells and colon tumour CC531 in Wag rats. The bimAb was produced by somatic hybridization of two mouse hybridomas, one producing monoclonal antibodies (mAb) against CD8 (IgG2b, OX8), and the other producing mAb against a CC531-associated antigen (IgG1, CC52). A bimAb-producing clone was selected by an enzyme-linked immunosorbent assay with CC531 tumour cells. BimAb were purified from ascitic fluid by protein A affinity chromatography. Each of five pooled peak fractions was analysed by flow cytometry for the presence of bimAb. Most bimAb were found in a fraction that was eluted at pH 4.5 from protein A. FPLC analysis of this fraction revealed that no parental antibodies were present. The OX8 x CC52 bimAb greatly increased conjugate formation in vitro between ALAK cells and CC531. Results of 51Cr-release assays with CC531 as target cells and ALAK cells as effector cells were not significantly different in the presence or in the absence of the bimAb. The methods we used here, a cell enzyme-linked immunosorbent assay and flow cytometry, are simple methods for development and purification of a bimAb when a functional selection method is not a priori available. The OX8 x CC52 bimAb we developed this way may increase in vivo tumour targeting of ALAK cells and thus augment antitumour effect in vivo.
- Subjects :
- Animals
Antibodies, Monoclonal biosynthesis
Antibodies, Monoclonal isolation & purification
CD8 Antigens immunology
Cell Line
Chromatography, High Pressure Liquid
Flow Cytometry
Male
Rats
Rats, Inbred Strains
Staphylococcal Protein A analysis
Tumor Cells, Cultured
Antibodies, Monoclonal immunology
Antibody Specificity
Colonic Neoplasms immunology
Cytotoxicity Tests, Immunologic
Killer Cells, Lymphokine-Activated immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0340-7004
- Volume :
- 36
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Cancer immunology, immunotherapy : CII
- Publication Type :
- Academic Journal
- Accession number :
- 8500112
- Full Text :
- https://doi.org/10.1007/BF01742257