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© Pierre Lafaye
Astrocytes marqués par des anticorps VHH anti-GFAP. Des anticorps d'alpagas dirigés contre une protéine spécifique des astrocytes, la GFAP (Glial Fibrillary Acidic Protein), ont été obtenus à partir de camélidés immunisés. La partie VHH (partie de l'anticorps qui reconnaît l'antigène) a été exprimée sous forme recombinante chez Escherichia coli.
Scientific Fields
Diseases
Organisms
Applications
Technique
Starting Date
26
Mar 2015
Status
Ongoing
Members
5
Structures
1

About

What we do

Phage display technology enables the selection from repertoires of VHH displayed on the surface of filamentous bacteriophage. Llamas or Alpacas are immunized with antigens provided by the users; once the serum shows an adequate immune response, blood lymphocytes are purified and the VHH genes population is isolated from lymphocytes then converted to phage-display format using PCR. Subsequent transformations usually yield libraries of 107 to 109 clones, each clone corresponding to a specific VHH. The library is panned against the antigen to select specific VHH. These VHH are then expressed in Escherichia coli and their characteristics are analyzed (purity, affinity, specificity).

The affinity of selected nanobodies are performed by biolayer interferometry (Octet HTX) and epitope mapping by HDX-Ms (collaboration with PF RMNbio).

Our team carries out support tasks and develops new protocols for immunization (DNA or cell-based immunization) and for site-specific labeling of nanobodies.

Access Protocol

Regular open desks are organized with other core facilities involved in protein science (from production to structure, dynamics and biophysics) to make a first contact with the team.

Alternatively, a request for support starts by sending a mail (anticorps@pasteur.fr). A meeting is organized to analyse the feasibility of the project and the needs of the applicant. Then a form has to be filled and submitted as a project.

Formulaire  PFIA  nanobodies

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