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AMB Express
Fig. 1 | AMB Express

Fig. 1

From: Preparation and identification of a single domain antibody specific for adenovirus vectors and its application to the immunoaffinity purification of adenoviruses

Fig. 1

Schematic diagram of the construction and screening of a single-domain phage display library. The Bactrian camel was immunized with adenoviruses rAd26 and ChAd63, and the blood was collected to isolate peripheral blood mononuclear cells (PBMCs). Total RNA was extracted from PBMCs and reverse transcribed into cDNA. The VHH gene fragments were amplified using PCR. In the first round of PCR, a fragment of approximately 600 bp from the leader sequence to the CH2 domain was amplified from the cDNA. The full-length VHH gene from FR1 to FR4 was amplified using P3 and P4 primers in the second round of PCR. The PCR-amplified product was digested using the restriction enzymes NcoI and NotI. The VHH gene was ligated into the pMECS plasmid and transformed into E. coli TG1. After rescue by the M13k07 helper phage, the VHH gene was displayed on the phage surface, and the specific phages were enriched and selected using phage ELISA

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