Thermo-stability and antitumor activity on colon cancer cell lines of monoclonal anti-CEA antibody-saporin immunotoxin

J Korean Med Sci. 1992 Jun;7(2):128-35. doi: 10.3346/jkms.1992.7.2.128.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / therapeutic use*
  • Antineoplastic Agents, Phytogenic / administration & dosage
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Carcinoembryonic Antigen / biosynthesis
  • Carcinoembryonic Antigen / immunology*
  • Colonic Neoplasms / pathology*
  • Colonic Neoplasms / therapy
  • Hot Temperature
  • Humans
  • Immunotoxins / therapeutic use*
  • N-Glycosyl Hydrolases*
  • Plant Proteins / administration & dosage
  • Plant Proteins / pharmacology*
  • Ribosome Inactivating Proteins, Type 1
  • Saporins
  • Tumor Cells, Cultured / drug effects

Substances

  • Antibodies, Monoclonal
  • Antineoplastic Agents, Phytogenic
  • Carcinoembryonic Antigen
  • Immunotoxins
  • Plant Proteins
  • Ribosome Inactivating Proteins, Type 1
  • N-Glycosyl Hydrolases
  • Saporins