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Single-chain bispecific antibody (scBsAb) isone of the promising genetic engineering antibody formats for clinical application. But the effects of interlinker se-quences on the biological properties of bispecific single-chain antibodies have not been studied in detail. Three interlinker sequences were designed and synthesized, and denominated as Fc, HSA, 205Cu001f, respectively. Universal vectors with these different interlinker sequences for scBsAb expression in E.coli were constructed. A model scBsAb based on a reshaped single-chain antibody (scFv) against human CD3 and a scFv directed against human ovarian carcinoma were generated and expressed in E. coli. The results of SDS-PAGE andWestern blot showed that the different interlinker sequences did not affect the expression level of scBsAb. However, asdemonstrated by ELISA and pharmacokinetics studiesperformed in mice, scBsAbs with different interlinker se-quences had difference in the antigen-binding activities and terminal half-life time (T1/2b) in vivo, the interlinker HSAcould remarkably prolong the retention time of scBsAb inblood. These results indicated that the peptide sequence ofinterlinker could affect important biological properties ofscBsAb, such as antigen-binding properties and stability in vivo. So, selection of an appropriate interlinker sequence is very important for scBsAb construction. Optimal interlinker can bring scBsAb biological properties more suitable forclinical application.
Single-chain bispecific antibodies (scBsAb) isone of the promising genetic engineering antibody formats for clinical application. But the effects of interlinker se-quences on the biological properties of bispecific single-chain antibodies have not been studied in detail. Three interlinker sequences were designed A synthesized scBsAb based on a reshaped single-chain antibody (scFv) against human CD3 and a scFv directed against human ovarian carcinoma were generated and expressed in E. coli. The results of SDS-PAGE and Western blot showed that the different interlinker sequences did not affect the expression level of scBsAb. However, as demonstrated by ELISA and pharmacokinetics studies performed in mice , scBsAbs with different interlinker se-quences had difference in the antigen-binding activities and terminal half-li These results indicated that the peptide sequence ofinterlinker could affect important biological properties of scBsAb, such as antigen-binding properties and stability in vivo. So, selection of an appropriate interlinker sequence is very important for scBsAb construction. Optimal interlinker can bring scBsAb biological properties more suitable for clinical applications.