Discuss the factors involved in developing an immunoassay

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Reference no: EM132342124

Analytical Chemistry III

GENERAL OBJECTIVES:

1. Understand the principles, design, operation and applications of immunoassays

2. Understand the basic principles and applications of automation in the laboratory

3. Understand the general principles, operation and applications of electroanalytical methods.

1.1. Understand the principles of immunoassays

1.2. Discuss the types of labels used in immunoassays: radiolabels, enzymes, fluorescence.

1.3. Discuss the different separation techniques used to separate bound analyte from free: dextran-coated charcoal, second antibody, immobilisation

1.4. Understand the practical aspects of immunoassays including: preparation of hapten-carrier conjugates, immunisation, antibody detection, antibody titres, alibration, matrix effects

1.5. Discuss the shape and precision of standard calibration curves, including precision profiles

1.6. Discuss the advantages and disadvantages of immunoassays in terms of time, sensitivity, selectivity etc.

1.7. Discuss the factors involved in developing an immunoassay

1.8. Discuss the use of affinity chromatography as an immunoassay.

1.9. Discuss immobilisation and elution techniques used with affinity chromatography

1.10. Briefly discuss possible future directions for immunoassay

2.1. Discuss the processes occurring in an analysis that may have automation possibility

2.2. Discuss the difference between discrete analysers and continuous flow analysers

2.3. Understand the principles of flow injection analysis (FIA)

2.4. Draw a simple schematic of an FIA system

2.5. Discuss the effects of convection and diffusion on the concentration profile of analytes

2.6. Discuss applications of FIA including limited-dispersion and medium-dispersion applications, stopped-flow methods and flowinjection titrations

2.7. Discuss principles and applications of automatic samplers

2.8. Discuss the use of laboratory robots for sample preparation

2.9. Discuss the advantages and disadvantages of using automated systems for analysis

3.1 Draw a two-electrode cell for use in potentiometry

3.2 Discuss the basic principles of ion selective electrodes

3.3 Identify the terms in the Nernst equation

3.4 Describe the relationship between activity and concentration

3.5 Discuss the use of Total Ionic Strength Adjustment Buffer so that concentration is equivalent to activity

3.6 Discuss the effects of interfering ions using the potentiometric selectivity coefficient and the Nickolsky-Eisenmann equation

3.7 Calculate the percentage error of the ISE due to interference

3.8 Discuss the types of ISE with examples: glass membrane (pH); solid state membrane; ion exchange and liquid membrane

3.9 Discuss the use of standard additions and working curves to calibrate ISEs

3.10 Calculate the concentration of samples using the calibration methods in 3.8 above

3.11 Discuss the use of potentiometry in titration

3.12 Discuss the principles of amperometry and amperometric titration

3.13 Draw a diagram of the equipment used for an amperometric titration

3.14 Discuss the advantages of membrane and membrane-covered electrodes with examples

3.15 Discuss the use of modified electrodes with examples

3.16 Discuss the principles of voltammetry

3.17 Draw a schematic of a system for potentiostatic three-electrode linear-scan voltammetry

3.18 Discuss the principles of the electric double-layer

3.19 Describe the shape of the voltammetric curve for reversible and irreversible reactions

3.20 Understand the effect of charging current on the measurement and how to compensate for this

3.21 Explain the term diffusion current

3.22 List factors that affect the diffusion current

3.23 Understand the terms peak potential, half-wave potential, residual current

3.24 Discuss the use of square wave voltammetry and describe the waveform used

3.25 Discuss the principles of stripping voltammetry

3.26 Understand the principles of the electrodeposition step

3.27 Understand the difference between anodic, cathodic and adsorptive stripping voltammetry

3.28 Discuss the advantages of using stripping voltammetry

3.29 Explain the differences between using the peak height and peak area for calculation of a concentration.

3.30 Calculate the concentration of a sample, using working curve and standard addition methods, analysed using stripping voltammetry (peak height and peak area)

3.31 Discuss the principles of polarography

3.32 Understand the terms in the Ilkovic equation and how this affects diffusion current

3.33 Discuss the two main types of pulse polarography techniques: differential pulse polarography and square-wave polarography

3.34 Discuss the organic and inorganic applications of polarography

3.35 Discuss the use of hydrodynamic electrodes

3.36 Discuss the types of hydrodynamic electrode including rotating disk (RDE), wall jet, dropping mercury electrode (DME), tube and channel

3.37 Draw schematic profiles of streamlines at these electrodes

3.38 Understand the relationship between the limiting current and diffusion layer thickness using the equation

lL=nFAD/δ

3.39 Describe the characteristics of the DME

3.40 Draw a diagram of the apparatus for a DME

3.41 Understand the terms used in the Cottrell equation

3.42 Discuss the advantages and disadvantages of the DME

3.43 Discuss applications of the DME

3.44 Discuss the use of RDEs to investigate the kinetics of reactions

3.45 Discuss the types of double hydrodynamic electrodes including rotating ring disk electrode (RRDE), wall jet ring-disc electrode (WJRDE) and the tube/channel double electrode (TDE/CDE).

3.46 Discuss the use of double electrodes to investigate electron transfer

3.47 Discuss the principles of microelectrodes

3.48 Understand the difference in diffusion to a microelectrode when compared with a macroelectrode

3.49 Compare the voltammetric response of a microelectrode with that of a standard macroelectrode

3.50 Discuss the types of microelectrode configuration including disk, cylindrical, band, and ring.

3.51 Discuss factors involved with the construction of microelectrodes

Reference no: EM132342124

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