Premature antibodies with rapid reaction kinetics and their characterization for diagnostic applications

Kim, Dong‐Hyung; Seo, Sung Chul; Paek, Sanghyun; Lim, Guei-Sam; Paek, Se‐Hwan

doi:10.1016/j.ab.2011.09.006

Cited by 22 publications

(29 citation statements)

References 33 publications

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“…The extracted maximum current change was about 1.91µA, which is very close to the experimental observed one, 1.90µA. The extracted dissociation constant was obtained as 1.37x10 -9 M, which is also in the reasonable range for most IGg antibody-antigen complexes (3,(9)(10). The Langmuir equation shown in equation [3] can be rearranged as the following expression.…”

Section: Resultssupporting

confidence: 80%

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Study of Protein-Peptide Binding Affinity Using AlGaN/GaN High Electron Mobility Transistors

et al. 2013

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Antibody-immobilized AlGaN/GaN high electron mobility transistors (HEMTs) were used for detecting peptides and revealing the number of binding sites of the antibody and the dissociation constants. Two binding sites were found on this antibody for the peptide and the dissociation constants of the antibody-peptide complex were extracted from a two-binding-site model. The estimated dissociation constants are consistent with the detection limit of the sensor showing the important role of the dissociation constants in the performance of the sensors. The results demonstrate that the AlGaN/GaN HEMTs cannot only be used for biosensors, but also can be used for the biological affinity study.

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Section: Resultssupporting

confidence: 80%

“…The correlation coefficient, r 2 is 0.9926. These two binding constants are in a reasonable range of regular antibody-antigen binding constants (3,10). The antigen, the peptide, consisting of only 20 amino acids is pretty small and the antibody is allowed to bind two antigens on its two binding sites.…”

Section: Resultsmentioning

confidence: 98%

Study of Protein-Peptide Binding Affinity Using AlGaN/GaN High Electron Mobility Transistors

et al. 2013

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“…Nevertheless, the purified Fab moiety exhibited the same binding pattern with rapid reaction kinetics to the intact antibody. 13) We expect that the purified Fab moiety could be used to develop myoglobin detection methods, leading to opportunities in a diagnostic sensor chip for a continuous monitoring system.…”

Section: Discussionmentioning

confidence: 99%

“…13) The prepared IgGs (7 mg/mL, 28 mg, 4 mL) were dialyzed against 1× PBS buffer for 4 h at 4°C. Papain (15 mg/mL) (Sigma, P3125) was diluted in 4 mL digestion buffer (10 mM PBS, 20 mM EDTA, 10 mM cysteine_HCl (Thermo, 44889), adjusted pH 7.4).…”

Section: Fab Fragmentation and Purificationmentioning

confidence: 99%

Purification and characterization of Fab fragments with rapid reaction kinetics against myoglobin

Song

Kim

Park

et al. 2015

Bioscience, Biotechnology, and Biochemistry

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Myoglobin is an early biomarker for acute myocardial infarction. Recently, we isolated the antibody IgG-Myo2-7ds, which exhibits unique rapid reaction kinetics toward human myoglobin antigen. Antibodies with rapid dissociation kinetics are thought to be premature IgG forms that are produced during the early stage of in vivo immunization. In the present study, we identified the epitope region of the IgG-Myo2-7ds antibody to be the C-terminal region of myoglobin, which corresponds to 144–154 aa. The Fab fragment was directly purified by papain cleavage and protein G affinity chromatography and demonstrated kinetics of an association constant of 4.02 × 104 M−1 s−1 and a dissociation constant of 2.28 × 10−2 s−1, which retained the unique reaction kinetics of intact IgG-Myo2-7ds antibodies. Because a rapid dissociation antibody can be utilized for antibody recycling, the results from this study would provide a platform for the development of antibody engineering in potential diagnostic areas such as a continuous monitoring system for heart disease.

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“…The bandgap difference between AlGaN and GaN form two dimensional electron gas in the heterojunction interface without intentional doping. The mobility will increase since the electron can move quickly without colliding with any impurities or dopant ions [2][3][4]. They al so have s o m e characteristic features, such as good electrical property, chemically stable, biocompatible, thermal stability, low power consumption and non-bio toxicity.…”

Section: Introductionmentioning

confidence: 99%

Direct Detection of NT- Pro BNP As a Cardiac Biomarker Using High Electron Mobility Transistors in Physiological Salt Environment

et al. 2016

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Cardiovascular disease (CVD) is one of the major causes to claim millions of lives all across the globe every year. While the chances of CVD increase with age, it can also occur in any age group of people, both male and female. Despite all efforts and tremendous accomplishments in the precaution and treatment of CVD, its detection at an early stage is still a big challenge for scientists in this present modern world. Here, NT-pro BNP plays a crucial role in acute congestive heart failure since it is biologically inactive, binds better with its receptors and has a longer half-life than any other, making it an ideal biomarker. The detection method is based upon the ligand-receptor interaction, also called as immunoassay, which can be successfully detected with the use of High Electron Mobility Transistors (HEMTs) with short duration pulse applied at the gate electrode. With its unique characteristic features like good electrical property, chemical stability, biocompatibility, thermal stability and low power consumption HEMT sensors also have been used in several other applications like ion-sensing, pH sensing, gas sensing, etc. Therefore, the detection method becomes simple, fast, convenient and cost effective.

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Premature antibodies with rapid reaction kinetics and their characterization for diagnostic applications

Cited by 22 publications

References 33 publications

Study of Protein-Peptide Binding Affinity Using AlGaN/GaN High Electron Mobility Transistors

Study of Protein-Peptide Binding Affinity Using AlGaN/GaN High Electron Mobility Transistors

Purification and characterization of Fab fragments with rapid reaction kinetics against myoglobin

Direct Detection of NT- Pro BNP As a Cardiac Biomarker Using High Electron Mobility Transistors in Physiological Salt Environment

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