scFv multimers of the anti-neuraminidase antibody NC10:: length of the linker between VH and VL domains dictates precisely the transition between diabodies and triabodies
scFv multimers of the anti-neuraminidase antibody NC10:: length of the linker between VH and VL domains dictates precisely the transition between diabodies and triabodies
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DOI：
10.1093/protein/12.7.597
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发表时间：
1999-07-01
期刊：
影响因子：
--
通讯作者：
Hudson, PJ
中科院分区：
文献类型：
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作者：
Atwell, JL;Breheney, KA;Hudson, PJ
单链Fv抗体片段（scFv）掺入多肽接头以将V-H和V-L结构域拴在一起。具有5-12个残基长的接头的scFv分子不能折叠成功能性Fv结构域，而是与第二scFv分子缔合以形成二价二聚体（双抗体）。V-H和V-L结构域的直接连接进一步限制缔合并迫使三个scFv分子缔合成三价三聚体（三抗体）。我们已经通过构建一系列来自抗神经氨酸酶抗体NC 10的scFv，其中接头从一个到四个甘氨酸残基变化，确定了接头长度对scFv缔合的影响。含有三个和四个残基的接头的NC 10 scFv分子显示出对二聚体形成（双抗体）的强烈偏好，而一个或两个甘氨酸残基的接头长度阻止双抗体的形成并指导scFv缔合成三聚体（三抗体）。数据表明在接头长度从三个减少到两个甘氨酸残基时，从二聚体（双抗体）到三聚体（三抗体）的相对严格的转变。建模研究与作为与双抗体形成相容的最小接头长度的三个残基一致。NC 10 scFv多聚体和抗独特型Fab'之间形成的复合物的电子显微镜图像显示，二聚体对于抗原结合是二价的，三聚体是三价的。
Single-chain Fv antibody fragments (scFvs) incorporate a polypeptide linker to tether the V-H and V-L domains together. An scFv molecule with a linker 5-12 residues long cannot fold into a functional Fv domain and instead associates with a second scFv molecule to form a bivalent dimer (diabody). Direct ligation of V-H and V-L domains further restricts association and forces three scFv molecules to associate into a trivalent trimer (triabody). We have defined the effect of linker length on scFv association by constructing a series of scf;vs from anti-neuraminidase antibody NC10 in which the linker varied from one to four glycine residues. NC10 scFv molecules containing linkers of three and four residues showed a strong preference for dimer formation (diabodies), whereas a linker length of one or two glycine residues prevented the formation of diabodies and directed scFv association into trimers (triabodies). The data suggest a relatively strict transition from dimer (diabody) to trimer (triabody) upon reduction of the linker length from three to two glycine residues. Modelling studies are consistent with three residues as the minimum linker length compatible with diabody formation. Electron microscope images of complexes formed between the NC10 scFv multimers and an anti-idiotype Fab' showed that the dimer was bivalent for antigen binding and the trimer was trivalent.