Expression of a bispecific dsFv-dsFv′ antibody fragment in Escherichia coli
Expression of a bispecific dsFv-dsFv′ antibody fragment in Escherichia coli
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DOI：
10.1093/protein/13.10.725
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发表时间：
2000-10-01
期刊：
影响因子：
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通讯作者：
Dübel, S
中科院分区：
文献类型：
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作者：
Schmiedl, A;Breitling, F;Dübel, S
将3条多肽链分泌到大肠杆菌周质中，获得了由两个不同的二硫键稳定的Fv抗体片段组成的双特异二硫键稳定的Fv抗体片段(dsFv-dsFv‘)。用固定化金属亲和层析富集dsFv-dsFv‘分子，再用阴离子交换层析进一步纯化。所构建的重组抗体保留了两种亲本抗原结合的特异性，并能够将两种不同的抗原交联。所描述的dsFv-dsFv‘设计对于体内治疗应用可能具有特别的价值，因为预期改善的稳定性与最小的免疫原性相结合。
A bispecific disulfide-stabilized Fv antibody fragment (dsFv-dsFv') consisting of two different disulfide-stabilized Fv antibody fragments connected by flexible linker peptides was produced by secretion of three polypeptide chains into the periplasm of Escherichia coli. The dsFv-dsFv' molecules were enriched by immobilized metal affinity chromatography and further purified by anion-exchange chromatography. The recombinant antibody constructs retained the two parental antigen binding specificities and were able to cross-link the two different antigens. The described dsFv-dsFv' design might be of particular value for therapeutic in vivo applications since improved stability is expected to be combined with minimal immunogenicity.