Application of a Fab fragment of monoclonal antibody specific to N-terminal pro-brain natriuretic peptide for the detection based on regeneration-free electrochemical immunosensor

Abstract

N-Terminal pro-brain natriuretic peptide (NT-proBNP) is an important marker for heart failure that reflects ventricular volume expansion, ventricular overload and the degree of cardiac injury. To establish a sensitive and simple detection method for it in serum, a regeneration-free immunosensor with novel Fab fragment monoclonal antibodies was prepared. The sensor detected NT-proBNP from 0.04 to 2.5 ng/ml with a limit of 0.03 ng/ml. The immunosensor is therefore a simple, cost-effective method to detect NT-proBNP and the proposed immunoassay system would enable other proteins to be detected and open new opportunities for protein diagnostics.

Critical overview on the application of sensors and biosensors for clinical analysis

“…Zhuo et al [90] reported a better LOD (0.006 ng mL −1 ) and a wide linear range (0.02-100 ng mL −1 ) than Yi et al [91] did, reporting an LOD of 30 pg mL −1 and a linear range of 0.04-2.5 ng mL −1 . The main advantage of the immunosensor proposed by Yi et al [91] is the simple regeneration procedure, which includes washing the magnet with the buffer to remove the avidin-functionalized magnetic nanoparticles, antigen and antibodies.…”

“…According to Esteban-Fernández de Ávila et al [89], the low cost, ease of automation and miniaturization of multiplexed immunosensors, as well as the use of disposable massproduced electrodes, make this approach a promising alternative tool for developing point-of-care devices for in situ clinical diagnosis. Another biomarker for predicting cardiac risk is NT-proBNP, with a key role in the natriuretic, diuretic and vasodilatory systems, as well as in the inhibition of the renin-angiotensin-aldosterone system and the sympathetic nervous system [91]. Zhuo et al [90] and Yi et al [91] also reported electrochemical immunosensors for the detection of NT-proBNP based on gold-MWCNT composites and avidin-functionalized magnetic nanoparticles, respectively ( Table 2).…”

“…Another biomarker for predicting cardiac risk is NT-proBNP, with a key role in the natriuretic, diuretic and vasodilatory systems, as well as in the inhibition of the renin-angiotensin-aldosterone system and the sympathetic nervous system [91]. Zhuo et al [90] and Yi et al [91] also reported electrochemical immunosensors for the detection of NT-proBNP based on gold-MWCNT composites and avidin-functionalized magnetic nanoparticles, respectively ( Table 2). Zhuo et al [90] reported a better LOD (0.006 ng mL −1 ) and a wide linear range (0.02-100 ng mL −1 ) than Yi et al [91] did, reporting an LOD of 30 pg mL −1 and a linear range of 0.04-2.5 ng mL −1 .…”

“…Zhuo et al [90] and Yi et al [91] also reported electrochemical immunosensors for the detection of NT-proBNP based on gold-MWCNT composites and avidin-functionalized magnetic nanoparticles, respectively ( Table 2). Zhuo et al [90] reported a better LOD (0.006 ng mL −1 ) and a wide linear range (0.02-100 ng mL −1 ) than Yi et al [91] did, reporting an LOD of 30 pg mL −1 and a linear range of 0.04-2.5 ng mL −1 . The main advantage of the immunosensor proposed by Yi et al [91] is the simple regeneration procedure, which includes washing the magnet with the buffer to remove the avidin-functionalized magnetic nanoparticles, antigen and antibodies.…”

Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures

“…In this study we aim to investigate the effects of host strain, culture medium and aeration conditions on the production and extracellular leakage of Fab fragments in shaken E. coli cultures by the example of Fabs binding specifically to N-terminal pro-brain natriuretic peptide (NTproBNP), an important diagnostic marker of heart failure that can be detected from serum by an immunoassay applying the anti-NTproBNP Fabs [19]. Three different culture media were compared, all of them containing complex nutrients, but differing in their primary carbon source as well as in induction strategy.…”