Accession Number : ADA312073

Title :   Selective Protein Toxin Destruction Using Hydrolyzing Catalytic Monoclonal Antibodies.

Descriptive Note : Final rept.,

Corporate Author : HEBREW UNIV JERUSALEM (ISRAEL)

Personal Author(s) : Green, Bernard S.

PDF Url : ADA312073

Report Date : JUN 1993

Pagination or Media Count : 68

Abstract : The problem of amide hydrolysis using catalytic antibodies was analyzed and attacked. A novel, direct screening for catalysis method was developed which allows detection of such antibodies in hybridoma supernatants. The method (catELISA) involves a solid-phase substrate and an anti-product ELISA assy; it was found to be general and highly effective using ester-hydrolyzing (anti-phosphonate hapten) antibodies. By employing autoimmune-prone mouse strains, vastly increased numbers of catalytic antibodies for ester hydrolysis were produced. All attempts using catELISA and these exotic mouse strains to obtain amide- or peptide-hydrolyzing antibodies by immunization with simple phosphonate haptens as transition state analogs failed. Therefore, the synthesis of new, more sophisticated transition state analogs as haptens was carried out. 2-Aminoalcohols, aminophosphinates, cyclic phosphinates and phosphonimides were pursued; haptens for hydrolysis of acyl-proline, phenylalanyl amides were prepared. The many hybridomas studied using these haptens failed to produce arnide-hydrolyzing catalysts. Our results emphasize the need for additional approaches to attack this challenging problem. However, the results do show that powerful antibodies can be produced, at least for ester bonds, and they have made available new technologies that may eventually afford sequence-specific catalytic antibodies.

Descriptors :   *MONOCLONAL ANTIBODIES, *HYDROLYSIS, *TOXINS AND ANTITOXINS, *PEPTIDE HYDROLASES, DETECTION, SYNTHESIS, ENZYMES, PROTEINS, DESTRUCTION, SUBSTRATES, ANTIBODIES, ANALOGS, BONDING, CATALYSIS, TRANSITIONS, AMIDES, NUMBERS, ESTERS, PHOSPHONATES, ANTIGENS, SOLID PHASES, HYBRIDOMAS, PHOSPHINATES.

Subject Categories : Biochemistry
      Medicine and Medical Research
      Toxicology

Distribution Statement : APPROVED FOR PUBLIC RELEASE