Nanobody (single domain antibody) production

The production of nanobody primarily required to select a camelid animal for immunization for about 5–10 weeks. The effector B cells are extracted from the plasma peripheral lymph blood of the immunized camel to isolate the total RNA and cDNA synthesis for sequencing. Phage display technology is used to screen the specific binding ability of the nanobody. Finally, the phage vector was transferred to the Nb expression system to express the Nb proteins. Even though the Nb production technology is relatively complicated, the application of phage display technology and the screening steps of Nb are being gradually optimized.

Figure  Schematic overview of nanobody generation. (A) Extracting a cDNA from peripheral blood obtained from a camelid after immunized. (B) Extracting Total mRNA and reversely transcribing into cDNA, then amplifying the VHH, the phage display vector and the VHH are digested by restriction enzymes, and the two fragments are connected. (C) Constructing natural or immune camelid-derived nanobody libraries. (D–F) Obtaining antigen-specific nanobody by multiple screening (adopted from Sun et al. 2021).

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