Enzyme-Based Fluorescent Biosensors and Their Environmental, Clinical and Industrial Applications

Abstract

The last two decades has seen a significant increase in interest in biosensors. According to the Web of Science database, the number of scientific publications over the past 20 years increased from 309 in 1993 up to 3,467 in 2013 (Fig. 1).The demand for developing new detection methods, increasing interest in visualization techniques [1][2][3], emphasis placed on new drug discovery programs [4], diagnostic test development [5][6][7], and attempts to explain cellular action mechanisms and to trace a cell's metabolism pathway [7,8], and the possibility of using biosensors in numerous fields of application, (for example: medicine [9-11], environmental protection [12-14, 16-17], food industry [18], and defense industry [19]) has results in an increasing number of research projects targeted at biosensor design and fabrication. Biosensors used in environmental monitoring measure the toxicity effect based on the detection of a chemical compound or compound group by selective recognition of a biomolecule in the receptor layer and then by detection of a signal after passing through the transducer layer [16].The application of fluorescence biosensors in environmental protection applications [20][21][22][23][24][25], medical diagnostics [26][27][28][29][30][31][32][33][34][35], and industries [36][37][38][39][40][41][42][43][44][45] also is growing.Pol. J. Environ. Stud. Vol. 24, No. 1 (2015) AbstractEnzyme-based fluorescence biosensors and their applications in environmental protection, medicine, and industry are described. Biosensors used in environmental protection measure toxicity effects. A chemical compound or group of compounds is detected by the recognition of molecules in the receptor layer and then by detecting a signal passing through the transducer layer. Biosensors are classified according to the transduction method. Special emphasis is placed on optical biosensors, especially fluorescent biosensors, and such measurement techniques as FRET (Fröster resonance energy transfer), FLIM (fluorescence lifetime imaging), FCS (fluorescence correlation spectroscopy), and changes in fluorescence intensity. The phenomenon of fluorescence in biosensors and the selection of appropriate methods are described. The use of enzymes in the receptor layer and enzyme classification according to its category and functions used for analyte detection are presented. The fluorescence properties of enzymes resulting from possessing such cofactors as flavin or heme (prosthetic) groups are discussed. Several methods for enzyme immobilization, namely entrapment, adsorption, covalent immobilization, cross linking, and affinity interaction are described, and the use of enzymatic fluorescence biosensors in the detection of analytes is presented.

Microbiological Biosensors Based on Luminescence

“…Doskonałym rozwiązaniem wydają się być biosensory, które powszechnie stosowane są w ochronie środowiska [1][2][3], medycynie [4][5][6] przemyśle spożywczym [7][8][9] lub przemyśle obronnym. W całej rodzinie biosensorów można wyróżnić mikrobiologiczne biosensory bazujące na zjawisku luminescencji, które ze względu na ich dobrą czułość, szybką reakcję na zanieczyszczenie oraz szerokie spektrum temperatury i zakres pH, w których mogą pracować, są coraz częściej stosowane.…”

The phenomenon of fluorescence in immunosensors.

“…Biosensors find wide applications in medical diagnostics, (Justino et al, 2010), food industry (Leonard et al, 2003), environmental protection (Kłos-Witkowska, 2015), arms industry (Bartoszcze, 2003), agricultural industry (Rana et al, 2010) and biopharmaceutical research (Vo-Dinh & Cullum, 2000). According to the report (Thusu, 2010), biosensors are used in 47 applications and an increasing number of potential applications is recorded every year.…”

Investigation of Biosensor Potential Component Stability Caused by Influence of External Condition

“…Therefore, the recent years show increased interest in development of such devices, both with respect to research and construction. Biosensors are extensively used in:  environment protection [11][12][13][14] where they are a tool to detect water contaminants, harmful gases and various odorants  food industry [15][16][17][18][19][20][21][22] where they are used to detect pesticides examination of product freshness, harmful toxins contained in food, assessment of fungal contamination in grain and detection of specific odorant molecules in nourishment  defense industry, where they are used to detect warfare agents [22]  in medicine, where they are extensively used for diagnostics [23][24][25][26][27][28] to detect markers in cancerous processes, concentration of sugar in blood, detect bacteria and viruses or early identification of diseases, infections and metabolic disorders [29]. Despite the extensive applications of biosensors, there are continuous attempts to improve the existing devices and develop a new generation of sensors.…”