Thermo-stability and antitumor activity on colon cancer cell lines of monoclonal anti-CEA antibody-saporin immunotoxin.
10.3346/jkms.1992.7.2.128
- Author:
Jiradej MANOSROI
1
;
Sabine von KLEIST
;
Aranya MANOSROI
;
Fritz GRUNERT
Author Information
1. Department of Pharmaceutical Technology, Faculty of Pharmacy, Chiang Mai University, Thailand.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Saporin;
Monoclonal anti-CEA antibody;
Immunotoxin;
DSC;
Colon cancer
- MeSH:
Antibodies, Monoclonal/*therapeutic use;
Antineoplastic Agents, Phytogenic/administration & dosage/*pharmacology;
Carcinoembryonic Antigen/biosynthesis/*immunology;
Colonic Neoplasms/*pathology/therapy;
Hot Temperature;
Humans;
Immunotoxins/*therapeutic use;
*N-Glycosyl Hydrolases;
Plant Proteins/administration & dosage/*pharmacology;
Ribosome Inactivating Proteins, Type 1;
Tumor Cells, Cultured/drug effects
- From:Journal of Korean Medical Science
1992;7(2):128-135
- CountryRepublic of Korea
- Language:English
-
Abstract:
Eight saporin peaks were obtained from the purification of seed extracts of Saponaria officinalis L. Saporin peak No. 6 (SAP-6) showed the highest activity in the inhibition of protein synthesis (98%) in an in vitro translation study. An immunotoxin (IT) was prepared from SAP-6 conjugated to a monoclonal anti-CEA antibody 26/5/1 (mab B) using N-succinimidyl pyridyl dithiopropionate (SPDP) and 2-iminothiolane as a cross linker. Under thermal stability study by a DSC (differential scanning calorimetry), the IT showed a denature temperature of 75 degrees C. In in vitro translation studies, the purified IT showed the same activity as SAP-6 at 10(-7) M and 10(-9) M protein concentration at 0, 30 and 60-min incubation effects with mab B and SAP-6 not conjugated at 24-hr incubation periods on human promyelocytic cell line HL 60 and on human colon adenocarcinoma cell lines which were SW 403, LoVo and LS 174 T. SAP-6, mab B and IT had no cytotoxic effect on HL-60. The IT showed a higher cytotoxic effect than SAP-6 in CEA-positive cell lines. The IT demonstrated the highest cytotoxic effect of 51% inhibition of control at 10(-7) M on the LS 174 T.
