Biosensor Slides
Biosensor Slides
BY SIMISOLA A. OGUNDUGBA
ABSTRACT
The purpose of this research is to shed light on the subject of biosensors and its use as a viable means of detecting various pollutants in the environment.
INTRODUCTION
There is a worldwide effort towards the development of bioanalytical devices that can be used for detection, quantification and monitoring of specific chemical species. Conventional analytical techniques (e.g. G.C., Ads. Spec. Ph. metry), although highly precise, suffer from the disadvantages of high cost, complex sample preparations, the need for trained personnel and the fact that they are mostly laboratory bound (cannot be done on-site).
Dioxins as an example:
Formed as a by-product of the combustion of chlorophenols (mostly biocides); recognized as one of the most poisonous synthetic chemicals. Its effects include alleged birth defects (teratogenicity), severe skin disorders, carcinogenicity, mutagenicity, immuneresponse retardation, and bio-accumulation in aquatic organisms and people. (Hileman, 1994; U.S. E.P.A. , 2005a).
Classification of biosensors
According to the transduction (sensing) method as electrochemical, optical, piezoelectric and thermal sensors ; OR According to the bio-recognition principle as immunochemical, enzymatic, whole-cell and DNA biosensors
PROCESS STEPS
STEP I
The determination of initial enzymatic activity (A) using optimum conditions (pH, temp., subst. conc.)
STEP II
STEP III
The measurement of the residual activity (A*) i.e. activity after exposure of the biosensor to the inhibitor).
DESIGN CONSIDERATIONS
1. An extensive background knowledge of the enzyme characteristics: taking into account the specificity of the enzyme Determining the optimum conditions for the enzyme (pH, temperature, etc.)
2. Immobilization technique
The enzyme is attached to the transducer surface (graphite, gold film) for stability and to put it in close proximity with the transducer.
(A) covalent binding; (B) adsorption; (C) gel entrapment; (D)cross-linking; (E)microencapsulation; (F) adsorption-cross-linking.
3. Transducer Choice
Any of the transduction method could be chosen (electrochemical, optical, piezo-electric, thermal). Electrochemical transducers are common since it can directly measure biochemical reaction properties in terms of current (I), voltage (v), or conductance (G). Various configurations of EC transducers are amperometric (I), potentiometric (V), conductometric (G), etc.
4. Enzyme-Inhibitor System.
The long-term function of enzyme-based biosensors depends on this. The inhibition can be either reversible or result in an irreversible inactivation of the enzyme. Reversible inhibition can either be competitive (inhibitor competes with active site of enzyme) OR uncompetitive (no competition with active site but competes with ES complex ).
6. REGENERATION
Strong inhibition of the enzymes can present a serious problem for practical applications by limiting the reuse of biosensors. Some chemical substances called regenerators can be used to treat enzymes after each exposure to inhibitors. This reduces ageing of the enzyme. Examples are NaF, pyridine-2-aldoxime methyliodide (2-PAM), 4-formylpyridinium bromide dioxime (TMB4).
Transducer Equation
I = ZFkmC*
where Z, F, Km are constants, C* is the is the bulk concentration of the product species and I (amperes) is the value of the detected electric current. Typical current rating are usually in the 10-6 - 10 -9
5. Validation.
Despite the practical advantages of biosensors, they should be comparable to conventional analytical systems in terms of reliability and sensitivity. Biosensor measurements have to be verified by comparing them with the results of conventional chemical analysis.
COMPARATIVE ADVANTAGES OF BIOSENSORS OVER CONVENTIONAL METHODS On-site applications; Minimal sample treatment; Fast response times (typically 5-10 mins.); Low cost; Portability; Ease of use
CONCLUSION
The field of biosensor technology holds many exciting analytical applications for environmental use. Biosensor applications is not aimed at eliminating conventional methods but to complement it.
Some References:
1. Sara Rodriguez-Mozaza, et al, Biosensors for Environmental Monitoring: A global perspective (2004). 2. Aziz Amine, et al, Enzyme inhibition-based biosensors for food safety and environmental monitoring, Biosensors and Bioelectronics 21 (2006) 1405 1423. 3. Gamal A. E. Mostafa, Electrochemical Biosensors for the Detection of Pesticides, The Open Electrochemistry Journal, (2010), 2, 22-42.