Hello! Does anyone have an experience of using a commercial ELISA kit, designed for IgG detection in plasma/serum, for the detection of nanobodies against the same target (e.g. SARS-CoV-2 Spike protein)? Are there any pitfalls to consider (e.g. changing detector antibody to allow its binding to a VHH)?
What exactly do you mean with "IgG VHH fragments"? IgG and Nanobodies have different structures and sequences, so it is highly unlikely that an IgG-specific ELISA will detect nanobodies. Unless you have fusion protein with a Fc-part of IgG and a nanobody binding domain. Even if, your ELISA will not give you an accurate value.
Thank you for your answer! We want to assess the binding affinity of nanobodies to the same antigen as the one immobilised on the plate of a commercially available ELISA kit, designed for IgG detection. Hence I have had a thought of using it with changing the detector antibody and posed a question here to know if anyone has tried that. We shall opt for developing a custom assay then!
As nanobodies are usually the product of genetic engineering and phage- or ribosome display selection, the are usually constructed with specific tags for selection, detection and purification, e.g. His-tag, Myc-tag or Flag-tag. Check with the provider of your nanobodies to see whether this is the case for the ones you are using. If this is the case, you can use commercial antibodies against those tags for detection or, in a sandwich ELISA format, for immobilization.
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