Selection of similar single domain antibodies from two immune VHH libraries obtained from two alpacas by using different selection methods

- VHHs against GFAP have been obtained by Ribosome display after immunization of one alpaca with GFAP.
- VHHs against GFAP have been obtained by phage display after immunization of another alpaca with a human brain extract containing GFAP.
- All the VHHs were closely related and one VHH was found to be strictly identical in both studies.

The two most used methods to select camelid single-domain antibody-fragments (VHHs) are: displaying their repertoires on the surface of filamentous bacteriophages (phage display) or linking them to ribosomes (ribosome display). In this study, we compared specific VHHs isolated from two different immune libraries coming from two different alpacas by using these two selection methods.Three anti-GFAP (glial fibrillary acidic protein) VHHs were derived from an immune library obtained by ribosome display after immunization of one alpaca with purified GFAP, a protein expressed by astroglial cells. In parallel, three other anti-GFAP VHHs were derived from an immune library by phage display after immunization of another alpaca with a human brain tissue extract containing GFAP. All the VHHs were closely related and one VHH was found to be strictly identical in both studies. This highlights the selection pressure exerted by the camelid immune system to shape the paratope of an antibody against a defined antigen.