Abstract：
:Human hemoglobin (Hb) is a biomarker of several diseases, and monitoring of Hb levels is required during emergent surgery. However, rapid and sensitive Hb detection methods are yet to be developed. The present study established a rapid, convenient, and highly sensitive detection method for Hb in human serum using a bivalent antibody-enzyme complex (AEC). AECs are promising sensing elements because of their ability to bind specific targets and their catalytic activity that produce signals. We recently reported a convenient and universal method to fabricate bivalent AECs with two antibody fragments, using the SpyCatcher/SpyTag system. The present study applied a bivalent AEC for highly sensitive and quantitative detection of human Hb. The bivalent anti-Hb AEC was successfully prepared by incubating both N- and C-terminus SpyCatcher-fused glucose dehydrogenase and SpyTag-fused anti-Hb single-chain variable fragments at 4 °C. As expected, the bivalent AEC for Hb with a multimeric structure showed higher affinity than the monovalent AEC, by means of avidity effects, unlike that for soluble epidermal growth factor receptor with a monomeric structure; this contributed to a great improvement in sensitivity. Finally, we established a rapid and wash-free homogeneous electrochemical detection system for Hb by integrating magnetic beads. The linear range of the system completely covered the clinically required Hb levels, even in human serum. This technology provides an ideal point-of-care test for Hb and other multimeric biomarkers.
journal_name
Talantajournal_title
Talantaauthors
Miura D,Kimura H,Tsugawa W,Ikebukuro K,Sode K,Asano Rdoi
10.1016/j.talanta.2021.122638keywords:
["Bivalent antibody-enzyme complex","Catcher\/tag system","Glucose dehydrogenase","Hemoglobin","Homogeneous electrochemical detection","Single-chain variable fragment"]subject
Has Abstractpub_date
2021-11-01 00:00:00pages
122638eissn
0039-9140issn
1873-3573pii
S0039-9140(21)00559-2journal_volume
234pub_type
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