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Do you need an affine antibody recognizing your cell surface receptor? Let us select a nanobody for you.
Hybrigenics can select nanobodies by phage display directly on living cells when in vitro selection is not possible e.g for receptors with transmembrane domains like GPCR's. Our hs2dAb library coding for 3.109 VHH nanobodies is expressed at the phage surface and incubated with the cell expressing the antigen of interest. After a washing step with cells that do not express the antigen, the elution of the specific nanobodies is performed and their enrichment is measured. 3 to 4 rounds of selection are performed.
Selected nanobodies are validated by immunofluorescence (IF). Each nanobody is tagged and expressed as an E.coli supernatant. Its colocalization with the antigen is detected with a secondary labeled anti-tag antibody by fluorescence microscopy.
Selected VHHs can also be validated by Phage FACS. After selection by phage display selection, cells are incubated with an anti-M13 antibody labelled with a fluorescent marker and sorted in a FACS. Fluorescence signal is compared between cells expressing the antigen of interest and cells which do not.