DIABODIES - SMALL BIVALENT AND BISPECIFIC ANTIBODY FRAGMENTS
DIABODIES - SMALL BIVALENT AND BISPECIFIC ANTIBODY FRAGMENTS
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DOI：
10.1073/pnas.90.14.6444
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发表时间：
1993-07-15
影响因子：
11.1
通讯作者：
WINTER, G
中科院分区：
文献类型：
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作者：
HOLLIGER, P;PROSPERO, T;WINTER, G
双价和双特异性抗体及其片段具有巨大的实际应用潜力。在这里，我们描述了带有两个抗原结合位点的小抗体片段的设计。所述片段包括与相同多肽链(V(H)-V(L))上的轻链可变区(V(L))相连的重链可变区(V(H))。通过使用太短而不能在同一链上的两个结构域之间配对的连接子，这些结构域被迫与另一个链的互补结构域配对，并创建两个抗原结合位点。二聚体的计算机图形模型表明，这两对结构域可以堆积在一起，而抗原结合部位指向相反的方向。二聚体抗体片段，或“双链抗体”，可以设计为二价或双特异性相互作用。从分别针对半抗原苯唑酮和鸡蛋溶菌酶的单抗NQ11.7.22(NQ11)和D1.3出发，构建了二价片段(V(H)NQ11-V(L)NQ11)2和(V(H)D1.3-V(L)D1.3)2和双特异性片段V(H)NQ11-V(L)D1.3和V(H)D1.3-V(L)NQ11。这些片段通过细菌分泌物表达，并被证明与半抗原和/或抗原特异性结合。5-残基和15=残基与亲本抗体的结合亲和力相似，但V(H)结构域直接连接到V(L)结构域的片段具有较慢的解离动力学和较高的对半抗原的亲和力。DIABADIES提供了一种在细菌中构建小的双价和双特异性抗体片段的现成方法。
Bivalent and bispecific antibodies and their fragments have immense potential for practical application. Here we describe the design of small antibody fragments with two antigen-binding sites. The fragments comprise a heavy-chain variable domain (V(H)) connected to a light-chain variable domain (V(L)) on the same polypeptide chain (V(H)-V(L)). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites. As indicated by a computer graphic model of the dimers, the two pairs of domains can pack together with the antigen-binding sites pointing in opposite directions. The dimeric antibody fragments, or ''diabodies,'' can be designed for bivalent or bispecific interactions. Starting from the monodonal antibodies NQ11.7.22 (NQ11) and D1.3 directed against the hapten phenyloxazolone and hen egg lysozyme, respectively, we built bivalent fragments (V(H)NQ11-V(L)NQ11)2 and (V(H)D1.3-V(L)D1.3)2 and bispecific fragments V(H)NQ11-V(L)D1.3 and V(H)D1.3-V(L)NQ11. The fragments were expressed by secretion from bacteria and shown to bind specifically to the hapten and/or antigen. Those with 5- and 15=residue linkers had similar binding affinities to the parent antibodies, but a fragment with the V(H) domain joined directly to the V(L) domain was found to have slower dissociation kinetics and an improved affinity for hapten. Diabodies offer a ready means of constructing small bivalent and bispecific antibody fragments in bacteria.