Accession Number:

ADB095321

Title:

Biological Synthesis of a Protein Analogue of Acetylcholinesterase: Monoclonal Anti-Idiotype Antibody Analogue of the Esteratic Site

Descriptive Note:

Annual rept. 15 May 1983-14 May 1984

Corporate Author:

JOHNS HOPKINS UNIV BALTIMORE MD SCHOOLOF MEDICINE

Personal Author(s):

Report Date:

1984-07-10

Pagination or Media Count:

21.0

Abstract:

The goal of this research during the first year of the contract was to develop a method for the purification of human erythrocyte acetylcholinesterase and to initiate the preparation and analysis of monoclonal antibodies. This was accomplished by the preparation of red blood cell membrane ghosts, enzyme solubilization with a non-ionic detergent, and enzyme purification by monoclonal antibody affinity chromatography. Sixty ml of packed red blood cells yielded a final fraction of 750 micrograms. approximately 20, 000-fold purified. The purified fraction contained a single protein of about 75, 000 daltons that was labeled with 3H-diisopropylfluorophosphate and gave a single peak during high-performance liquid chromatography on a TSK-SW3000 silica-enzyme for the preparation of monocloninesterase, anti-active site, and anti-idiotype monoclonal antibodies have been developed.

Subject Categories:

  • Biochemistry

Distribution Statement:

APPROVED FOR PUBLIC RELEASE