The first constant domain (CH1 and CL) of an antibody used as heterodimerization domain for bispecific miniantibodies

FEBS Letters

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 C_H1 domain of an IgG1 to the C-terminus of a single-chain Fv fragment (scFv-425), specific for the EGF receptor, and fusing the C_L 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 C_H1-C_L 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