Services
mmunization of a llama or alpaca with the target antigen (usually purified protein provided by the user, other immunization strategies can be discussed)
Serum ELISA to verify immune response against the target antigen
Generation of VHH phage libraries encoding the VHH repertoire of a blood sample
Enrichment of target specific VHHs by phage display (different selection strategies)
Identification of nanobody candidates by ELISA
Small scale production and purification of preliminary hits
Validation of custom-made nanobodies Binding assay in vitro
Procaryotic expression in the periplasma of E.coli (Nanobodies, Nanobody-Dimers)
Eukaryotic Expression in HEK6e cells (Nanobodies, FC-Fusion, Dimer, Multimers, BiTEs)
FC fusion: reconstitition of heavy chain antibody format. Different species (human, mouse, rabbit) and mutations (enhanced CDC, ADCC or silent)
Half life extension: Nanobody Dimer with one Nanobody directed against albumin
Dimers and Multimers: Homo-Dimers, Hetero, Dimers, different forms of multimers
Fusion proteins: Nanobodies fused to target protein, e.g. GFP, mCherry for imaging
Different Tags for IP, Staining, conjugation etc. (HA, Alfa, Avi, twinstrep, His, Sortase, ...)
Site specific conjugation by Sortase A: Enzymatic conjugation of any moiety with N-Term. GGG to LPETG tag (e.g. fluorochromes, biotin, drugs)
Site specific biotinylation by BirA: Higly efficient Biotinylation of Avi-Tag, in vitro or during eukaryotic co-expression of nanobody and BirA enzyme
NHS conjugation: Tag-independent modification af amino-residues by NHS-Ester formation. Broadly appicable to conjugate small molecules, beads, peptides, etc.
Binding on recombinant protein by ELISA
Binding on target expressing cells by FACS
Epitope Binning by ELISA
Epitope Binning by Biolayer Interferometry
Affinity estimation by ELISA
Affinity determination by Biolayer Interferometry
Address
Arrival by bus and train or map
From Bonn main station you can take the bus line 600 (direction Ippendorf Altenheim) and 601 (direction Venusberg) to the bus stop “Uniklinikum Nord, Uniklinikum Hauptpforte or Uniklinikum Süd”. Line 630 runs in the Bonn city area without stopping at the main station. Please use the night bus (N5) between midnight and 5:30 a.m.
The Core Facilities thank the German Research Foundation for continuous support.
