期刊
PROTEIN ENGINEERING DESIGN & SELECTION
卷 17, 期 4, 页码 315-323出版社
OXFORD UNIV PRESS
DOI: 10.1093/protein/gzh040
关键词
antibody fragments; biodistribution; breast cancer; HER2; minibody
资金
- NCI NIH HHS [CA 16042, CA 48780, CA 43904, CA 33572] Funding Source: Medline
An engineered antibody fragment (minibody; scFv-C(H)3gamma(1) dimer, M, 80 000) specific for carcinoembryonic antigen (CEA) has previously demonstrated excellent tumor targeting coupled with rapid clearance in vivo. In this study, variable (V) genes from the anti-p185(HER-2) 10H8 antibody were similarly assembled and expressed. Four constructs were made: first, the V genes were assembled in both orientations (V-L-linker-V-H and V-H-linker-V-L) as single chain Fvs (scFvs). Then each scFv was fused to the human IgG1 C(H)3 domain, either by a two amino acid linker (ValGlu) that resulted in a non-covalent, hingeless minibody, or by IgG1 hinge and a GlySer linker peptide to produce a covalent, hinge-minibody. The constructs, expressed in NS0 mouse myeloma cells at levels of 20-60 mg/l, demonstrated binding to the human p185(HER-2) overexpressing breast cancer cell line, MCF7/HER2. Binding affinities (K-D similar to 2-4 nM) were equivalent to that for the parental 10H8 mAb (K-D similar to 1.6 nM). Radioiodinated 10118 hinge-minibody was evaluated in athymic mice, bearing MCF7/HER2 xenografts. Maximum tumor uptake was 5.6 (+/-1.65)% injected dose/g (ID/g) at 12 h, which was lower than that of the anti-CEA minibody, whereas the blood clearance (beta-phase, 5.62 h) was similar. Thus, minibodies with different specificities display similar pharmacokinetics, while tumor uptake may vary depending on the antigen-antibody system.
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