Direct immobilization of gold-binding antibody fragments for immunosensor applications

Takahisa Ibii, Masaru Kaieda, Satoru Hatakeyama, Hidenori Shiotsuka, Hideki Watanabe, Mitsuo Umetsu, Izumi Kumagai, Takeshi Imamura

Research output: Contribution to journalArticlepeer-review

35 Citations (Scopus)


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 (KD ∼ 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 (KD) 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.

Original languageEnglish
Pages (from-to)4229-4235
Number of pages7
JournalAnalytical Chemistry
Issue number10
Publication statusPublished - 2010 May 15


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