How can we evaluate the affinity of an antibody towards a specific antigen if the antibody has been expressed on the membrane of E. coli?

05 June 2023 1 10K Report

Hello! We used plasmid pNeae2 to display nanobodies on the surface of E. coli DH10B-T1 R using the bacterial display methodology. Then we attempted to use this bacterium as a primary antibody in an indirect ELISA assay to test its affinity with a protein. If anyone has any suggestions regarding this methodology or references related to it, please let us know.

tags: nanobodies, single domain antibodies, VHH antibodies, indirect ELISA assay

Ahmed Thabet

Cytoplasmic expression of antibody fragments in E. coli often results in the production of aggregates within inclusion bodies. Antigen binding activity can be reconstituted after polypeptide refolding1,2 but recovery takes a hit in this process

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