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. 2023 Apr 19;15(1):2195517. doi: 10.1080/19420862.2023.2195517

Figure 3.

Crystal structures of several spFv molecules alone and some in complex with target proteins show stapling disulfide bonds form as designed. These images also show that orientation (LH or HL) or antigen binding does not impact the Fv structures.

Structures and comparison of various scFv/spFv domains. In all structures, VL is colored blue and VH is colored green. The linker segments are colored pink. (a) GLk1 spFv LH. (b) GLk1 spFv HL. (c) GLk2 spFv HL. (d) 2mfo-dFc electron density contoured at 1.5 σ about the CPPC staple motif and anchor points for Glk2 spFv HL. Circles in orange indicate the stapling disulfide density. (e) CAT2200b spFv HL. (f) unbound CAT2200b spFv HL compared with CAT2200a scFv LH bound to IL-17 (silver). (g) front and back views of unbound CAT2200b spFv HL compared with CAT2200a spFv LH bound to IL-17 (silver).