Phage display and production of recombinant single-domain antibodies

Antigen design
and camelid immunization

• Native or recombinant proteins, bioconjugated or derivatized haptens.
• Immunization.
• Purification of circulating lymphocytes (PBMC).
• Construction of a cDNA library.

Construction of a
phage display library

• Phage library expression carrying V_HH.
• Selection of clones with specificity for the antigen (panning).

Library
Screening

• Antigen affinity evaluation.
• Amino acid sequencing of V_HH candidates.

Affinity maturation and engineering for functionalization

Small-scale
production

• V_HH phage-free expression in E. coli.