Principles and Techniques of Biochemistry

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Master in Advance Nanoscience and Nanotechnology, Universitat Autònoma de Barcelona. 2019/2020

General Info

Schedule

Week 1: 12s (18-20 h); 13s (15-18 h)

Week 2: 19s (15-18 h)

Week 3: 25s (17-18 h)

Evaluation: not determined

 

Location:  CB/012

Assessment

The competences of this subject will be evaluated by means of The following  activities. 

  • First deliverable (cooperative, deadline 01/10/19).

Select commercial antibodies for the following targets (antigen)

– Salmonella (multideterminate, multivalent example)

-Glyphosate (hapten, unideterminate and univalent example) 

sources: CiteAb and Abcam

  • Second deliverable (deadline 01/10/19). 

Describe, step by step, an ELISA test for the detection of Listeria infection. It can be focused on the detection of antibodies against the bacteria or the bacteria.  Make an scheme to describe the format and add detail of the commercial antibodies selected to achieve this task.

Topics

Biomolecules and Biorecognition

Amino Acids, Peptides and Proteins.    Antibodies.   Enzymes.   Nucleic Acid Other affinity interactions in nature. Strept(avidin), Protein A and G. Aptamers  Biorecognition: Enzyme/Substrate. Antigen/antibody. Hybridization.

Separation methods of biomolecules

Electrophoresis of proteins and DNA. Gel Electrophoresis (GE). SDS-Polyacrylamide Gel Electrophoresis (SDS–PAGE). Applications of Electrophoresis. Diagnosis of Genetic (Inherited) Disease. Paternity and forensic testing. DNA Sequencing. Diagnosis of infection diseases. Chromatography. Liquid Chromatography for Bioanalysis. Reversed Phase Liquid Chromatography (RP-LC). Ion Exchange Chromatography (IEC). Size Exclusion Chromatography (SEC). Affinity Chromatography.

 

Production of bioreagents

Synthesis of short peptides and oligonucleotides. Production of monoclonal and polyclonal antibodies. Antibodies fragmentation. Synthesis the immunogens. Purification of proteins from nature sources or cultures.

 

Immunoassays

Classification. Antibody-Antigen Complex Formation. Assay Formats. Homogeneous and heterogeneous immunoassays. Labels in immunoassays: radioisotopes, fluorescence, chemiluminescent, enzymes. Enzyme Immunoassays: ELISA.     Rapid diagnostic tests: lateral flow. The pregnancy test.

DNA analysis and amplification

Hybridization reaction of DNA. DNA amplification: PCR (polymerase chain reaction). The Principle of PCR. Reagents for PCR.

Immobilization and labeling of biomolecules

Solid supports in bioanalytical chemistry. Immobilization strategies of biomolecules in solid supports.Non-specific adsorption. Blocking reagents. Labeling of biomolecules: enzymes, fluorophores, nanoparticles, QDs, biotin. Signal amplification strategies.

Bibliography

– David L. Nelson, Michel M. Cox. ‘Lehninger Principios de Bioquímica’. 4ª ed. Ediciones Omega. 2006.
– L. Stryer, Jeremy M. Berg, John L. Tymocxko. Bioquímica. 5 ª ed. Editorial Reverté. 2003.
– Mary K. Campbell, Shawn O. Farrell. Bioquímica. 4 ª ed. Editorial Thomsom. 2004.

 

Bioanalysis. Other suggested bibliography

– Bryan L. Williams, Keith Wilson. Principios y Técnicas de Bioquímica experimental. Ediciones Omega, 1981.
– J.M. García Segura, J.G. Gavilanes, A. Martínez del Pozo, F. Montero, M. Oñaderra, F. Vivanco. Técnicas instrumentales de análisis en Bioquímica. Editorial Síntesis, 2002.
– D.J. Holme, H. Peck. Analytical Biochemistry. 3rd edition. Prentice Hall, 1998.
– Rodney F. Boyer, Modern Experimental Biochemistry (Third Edition) Benjamin/Cummings, San Francisco, 2000.

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