| abstract |
The present application relates to the technical field of antibody library construction, in particular to a method for constructing a semi-synthetic nanobody library and a nanobody library. The method includes: using the cDNAs of CDR1, CDR2 and CDR3 as templates, performing a first amplification to obtain CDR1 respectively. , the base sequences of CDR2 and CDR3; the nucleotide sequences of FR1, FR2, FR3 and FR4 are connected with the base sequences of the CDR1, the CDR2 and the CDR3, and the second amplification is carried out to obtain the target gene fragment; connecting the target gene fragment with the first vector to obtain a second vector; after culturing the second vector, screening to obtain a target vector containing a nanobody library, wherein the base sequence connection relationship of the target gene fragment Includes: FR1‑CDR1‑FR2‑CDR2‑FR3‑CDR3‑FR4. A semi-synthetic nanobody library is constructed by using the antibody skeleton of FR1, FR2, FR3 and FR4 and synthesizing part of the CDR region, avoiding the need for existing nanobodies to be obtained by antigen immunization. Save cost and time. |