Corporate R&D Headquarters, Frontier Research Center, Canon Inc., 30-2, Shimomaruko 3-chome, Ohta-ku, Tokyo 146-8501, Japan.
Anal Chem. 2010 May 15;82(10):4229-35. doi: 10.1021/ac100557k.
A novel method that enables antibody fragments to be immobilized on a sensor substrate with a high binding capability using molecular recognition has been developed. Using genetic engineering, we fabricated bispecific recombinant antibody fragments, which consist of two kinds of antibody fragments: a gold antibody fragment and a target molecule antibody fragment. Surface plasmon resonance (SPR) analysis indicated that these gold-binding bispecific antibody fragments bind directly to the gold substrate with high affinity (K(D) approximately 10(-9) M). About 70% of the bispecific antibody fragments immobilized on the gold substrate retained their target protein-binding efficiency. The Sips isotherm was used to assess the heterogeneity in antibody affinity for the bispecific antibody fragments. The results showed that the immobilized bispecific antibody fragments exhibited an increased homogeneity of affinity (K(D)) to target molecules when compared with monospecific antibody fragments immobilized by conventional methods. The use of bispecific antibody fragments to directly immobilize antibody fragments on a solid-phase substrate offers a useful platform for immunosensor applications.
一种新方法，利用分子识别将抗体片段高亲和力地固定在传感器基质上，已被开发出来。我们使用基因工程制造了双特异性重组抗体片段，它由两种抗体片段组成：一种是金抗体片段，另一种是靶分子抗体片段。表面等离子体共振（SPR）分析表明，这些与金结合的双特异性抗体片段以高亲和力（K(D)约为 10(-9) M）直接与金基质结合。大约 70%固定在金基质上的双特异性抗体片段保留了其靶蛋白结合效率。Sips 等吸附等温线用于评估抗体对双特异性抗体片段亲和力的异质性。结果表明，与通过传统方法固定的单特异性抗体片段相比，固定在双特异性抗体片段上的抗体片段对靶分子的亲和力（K(D)）均匀性增加。使用双特异性抗体片段将抗体片段直接固定在固相基质上，为免疫传感器应用提供了一个有用的平台。
