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Title 

Construction of an antimyoglobin single-chain variable fragment with rapid reaction kinetics

Authors 

J H JangD H KimS H PaekEui-jeon WooY W Kim

Publisher 

Portland Press

Issue Date 

2016

Citation 

Biotechnology and Applied Biochemistry, vol. 63, no. 1, pp. 22-30

Keywords 

acute myocardial infactioncontinuous monitoringmyoglobinrapid reaction kineticssingle-chain variable fragment

Abstract 

Antibodies with rapid reaction kinetics (high association and dissociation rates), named reversible antibodies, are used to perform continuous monitoring of sensitive disease biomarkers. In cases of acute myocardial infarction (AMI), continuous monitoring and early diagnosis are important. Human myoglobin (Myo) is a useful biomarker for AMI during the early stage after the onset of symptoms. In this study, a single-chain variable fragment (scFv) specific to Myo was derived from an IgG antibody that has rapid reaction kinetics. Enzyme-linked immunosorbent assay revealed that recombinant scFv exhibited 3.8-fold reduced affinity compared with the parent IgG antibody based on the antibody concentration necessary for 50% of the maximum signal. The scFv retained the rapid reaction kinetic mode with average kon and koff of 2.63 × 105 M-1 Sec-1 and 3.25 × 10-3 Sec-1, respectively, which were reduced to 10- and 2.3-fold compared with those of the parent antibody. The equilibrium constant for the association of the scFv (KA = 8.09 × 107 M-1) was 4.6-fold lower than that of its parent IgG antibody. This scFv may be a starting point for further mutagenesis/kinetic and structural analyses providing valuable insight into the mechanism of reversible antibodies.

ISSN 

0885-4513

Link 

http://dx.doi.org/10.1002/bab.1349

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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