test_moissonnage_selune
type: Agent

Exploration of recombinantes proteins expression systems for the characterization of a catalytic antibody
type: Dataset
contact point: Organization
  fn: CCSD
keyword: renaturation
keyword: anticorps-catalytiques
keyword: cytoplasmic-expression
keyword: theses
keyword: refolding
keyword: spiengineering-sciences-physics
keyword: single-chain-variable-fragment-scfv
keyword: mammalian-expression
keyword: infoeu-reposemanticsdoctoralthesis
keyword: rps
keyword: expression-cytoplasmique
keyword: expression-mammalienne
keyword: proteines-recombinantes
keyword: catalytic-antibodies
landing page: https://theses.hal.science/tel-00913012
description: Catalytic antibodies are investigated in order to understand their role under physio-pathological situations. But they also appear to be revolutionary tools to perform studies at the interface between chemistry, biochemistry, biology and immunology. Consequently, the knowledge of structure–function relationships is of great interest. We explored two expression systems for the production of a model catalytic antibody displaying a beta-lactamase activity. The recombinant scFv fragment was produced in the prokaryotic expression system. scFv fragments are often described as proteins being laborious to produce. An efficient method was developed to produce large quantities of refolded soluble catalytic scFv. Whole catalytic antibody was also produced by exploiting eukaryotic expression system. Mammalian cells were used because they are able to retain the original protein folding, assembly and post-translational modifications. The secondary structure of the catalytic scFv has been analyzed by circular dichroism to ensure that the refolded scFv is consistent with a native scFv fold. The functionality of the catalytic scFv and whole catalytic antibody has been validated by two approaches: (1) development of enzyme-linked immunosorbant assay (ELISA) and surface plasmon resonance (SPR) approaches for testing that the binding characteristics of an inhibitory peptide have been retained, and (2) proof of the subtle catalytic properties conservation through the development of a new sensitive catalytic assay using a fluorogenic substrate. This will lead to consider potential biotechnological and therapeutic applications of catalytic antibodies.
identifier: NNT: 2013COMP2078
issued: 2026-05-07T23:54:09.906992
language: fr
modified: 2026-05-07T23:54:09.906997

Exploration of recombinantes proteins expression systems for the characterization of a catalytic antibody
type: Distribution
access URL: https://theses.hal.science/tel-00913012
format: HTML
issued: 2026-05-07T23:54:09.918633
modified: 2026-05-07T23:54:09.894155