Anti-CD40 single-domain antibody, and preparation method therefor and use thereof
The present invention relates to the technical field of molecular biology, and in particular, to an anti-CD40 nanobody, and a preparation method therefor and use thereof. The anti-CD40 nanobody provided by the present invention has three unique complementarity determining regions, namely CDR1, CDR2, and CDR3. The present invention further provides a sequence encoding the nanobody or its VHH chain, a corresponding expression vector, and a host cell capable of expressing the nanobody. The present invention includes immunizing an alpaca by constructing a CD40 antigen to obtain a PBMC of the alpaca and constructing a vector; screening an anti-CD40 antibody through phage display; and finally obtaining a highly sensitive and specifically binding antibody through functional assays and sequencing result analysis. The nanobody provided by the present invention can specifically recognize and bind to CD40.
1 . An anti-CD40 single-domain antibody, comprising framework regions and complementarity determining regions (CDRs), wherein the CDRs comprise CDR1, CDR2, and CDR3, respectively corresponding to SEQ ID NO. 42, SEQ ID NO. 20, and SEQ ID NO. 21 in sequence.
2 . The anti-CD40 single-domain antibody of claim 1 , wherein an amino acid sequence of the anti-CD40 single-domain antibody is SEQ ID NO. 29.
3 . The anti-CD40 single-domain antibody of claim 1 , wherein a nucleotide sequence encoding the amino acid sequence of the anti-CD40 single-domain antibody is SEQ ID NO. 37.
4 . A type I transmembrane protein assay kit, comprising the anti-CD40 single-domain antibody according to claim 1 and known concentration of CD40 protein.