FEBS Letters | |
The first constant domain (CH1 and CL) of an antibody used as heterodimerization domain for bispecific miniantibodies | |
Arndt, Katja M2  Müller, Kristian M2  Strittmatter, Wolfgang1  Plückthun, Andreas2  | |
[1] Merck KGaA, D-64271 Darmstadt, Germany;Biochemisches Institut, Universität Zürich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland | |
关键词: Bispecific antibody; scFv fragment; Tumor targeting; CD2; EGF receptor; Ab; antibody; 425; antibody binding to EGF-R; EGF-R; epidermal growth factor receptor (extracellular domain); ELISA; enzyme linked immunosorbent assay; IMAC; immobilized metal-ion affinity chromatography; M1; antibody binding to CD2; PCR; polymerase chain reaction; PDB; protein data bank; scFv; single-chain Fv antibody fragment; SDS sodium dodecyl sulfate; SPR; surface plasmon resonance; | |
DOI : 10.1016/S0014-5793(98)00021-0 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
Bispecific miniantibodies were constructed by genetically fusing the CH1 domain of an IgG1 to the C-terminus of a single-chain Fv fragment (scFv-425), specific for the EGF receptor, and fusing the CL domain of a kappa light chain to the C-terminus of a scFv specific for CD2 (scFv-M1). An efficient dicistronic gene arrangement for functional expression in Escherichia coli was constructed. Immunoblots demonstrated correct domain assembly and the formation of the natural CH1-CL disulfide bridge. Gel filtration confirmed the correct size, sandwich ELISAs demonstrated bispecific functionality, and SPR biosensor measurements determined binding to EGF-R in comparison to bivalent constructs. Bispecific anti-EGF-R/anti-CD2 miniantibodies are candidates for the immunotherapy of cancer.
【 授权许可】
Unknown
【 预 览 】
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