Expression, Production, and Characterization of Recombinant Human Acetylcholinesterase and Its Mutants.

Abstract

Human cell line clones secreting high levels of fully active and soluble recombinant human acetylcholinesterase (rHuAChE) were developed by genetic engineering techniques. Some of the obtained clones produce as much as 60 units/10 6 cells/24 hours which is equivalent to 10 pg enzyme/cell/24 hour. Reaching such high production levels of rHuAChE from mammalian cells was one of the major goals of this contract. Development of biotechnological procedures, which will allow in the future to scale up the production of rHuAChE from these clones, has already been initiated. The expression vector systems that were developed were also used used to generate and study mutants of rHuAChE modified at the active center (Ser-203, His-432, his-447 mutants) or modified in ability to oligomerize (Cys-580 mutant) and produce active monomers of the enzyme.

Document Details

Document Type
Technical Report
Publication Date
Apr 01, 1991
Accession Number
ADB158891

Entities

People

  • Avigdor Shafferman

Organizations

  • Israel Institute for Biological Research

Tags

DTIC Thesaurus Topics

  • Acetylcholinesterases
  • Cell Line
  • Cells
  • Clones
  • Contracts
  • Engineering
  • Enzymes
  • Genetic Engineering
  • Production
  • Production Engineering

Fields of Study

  • Biology
  • Engineering

Readers

  • Molecular and Cellular Biochemistry
  • Neurotoxicology
  • Oncology (Cancer Research).

Technology Areas

  • Biotechnology