Grade in Chemistry, Universitat Autònoma de Barcelona. 19/20
Bioanalytical Chemistry arises as a result of the convergence of Analytical Chemistry and Biochemistry and addresses the design and application of novel specific and sensitive analytical techniques. The term Bioanalytical Chemistry involves not only the resolution of biological samples or bioanalytes, but also to the use of the biological reaction and the biomolecular reagent to obtain analytical information. This approach is influencing decisively in the development of new bioanalytical methodologies, with several advantages over traditional analytical methods. A main issue of classical Analytical Chemistry, the selectivity, can be solved by the integration of biological reagents, providing specificity. Furthermore, the biorecognition may simplify the analytical procedure by avoiding complex treatments of the sample. On the other hand, the integration of biological origin allows to improve the limits of detection.
Currently, the methods used in Bioanalytical Chemistry include, beside the classic analytical instrumental methods, such as chromatography and mass spectrometry, other methods derived from molecular biology, including PCR (polymerase chain reaction), enzymatic or immunological methods.
Problems and Exercises from 21/02 to 27/03 in C3/015, and from 27/03 in C3/019.
Lectures: from 11/02 to 27/03 in C3/015, and from 27/03 in C3/012
The competences of this subject will be evaluated by means of:
a) Middle term test (individual assessment), including the 1st part of the subject. 35% of the final mark. 20/04/2020 from 9 a.m. to 12 p.m.
b) Final term test (individual assessment), including the 2nd part of the subject. 35% of the final mark. 16/06/2020 from 9 to 12 p.m.
c) If a student fails in any of the two tests (mark below 5.0), there will be a final exam including the whole subject (individual assessment). 70% in the final mark. 30/06/2020 from 16 to 19 p.m.
d) Cooperative and collaborative activities. 30% of the final marks.
For more details about the assessment, download the General Guidelines
Problems and exercises
A password is required to download the files
Introduction to Bioanalytical Chemistry. Safety and risks in the biochemistry laboratory. The bioanalytical methodology. Obtaining samples. Qualitative (screening) assays vs. quantitative Assays. Data fitting and statistical treatment. Validation of bioanalytical methods. Sensitivity and specificity. Matrix effect. Interferences.
Structure of biomolecules and Biorecognition. Amino Acids, Peptides and Proteins. Antibodies. Enzymes. Nucleic Acids. Biorecognition: Enzyme/Substrate. Antigen/antibody. Hybridization. Other affinity interactions in nature. Strept(avidin), Protein A and G. Aptamers. Biomimetic recognition. The importance of water in biorecognition. Biological buffers.
Separation methods of biomolecules. Electrophoresis of proteins and DNA. Gel Electrophoresis (GE). SDS Polyacrylamide Gel Electrophoresis (SDS–PAGE). Southern, Northern and Western blot. Applications of Electrophoresis. Diagnosis of Genetic (Inherited) Disease. Paternity and forensic testing. DNA Sequencing. Diagnosis of infection diseases.
Chromatography. Introduction Part I and Part II. Liquid Chromatography for Bioanalysis. Reversed Phase Liquid Chromatography (RP-LC). Ion Exchange Chromatography (IEC). Size Exclusion Chromatography (SEC). Affinity Chromatography. Protein Sequencing. The special case of protein purification from natural sources
Instrumental techniques for the detection of biomolecules.Spectrophotometry and Fluorimetry. UV-VIS, turbidimetry, X-rays. Mass spectrometry for biomolecules. Genomics, proteomics and metabolomics.
Enzymatic analysis. Enzymes in bioanalytical chemistry. Enzymatic kinetics. Examples of reactions catalyzed by enzymes. Enzymatic inhibitors. Quantification of enzymes and their substrates. Clinical Examples. Creatinin. Glucose. Uric Acid. Urea.Colesterol. Aspartate aminotransferase (GOT or AST). Alanine aminotransferase (ALT or GPT). γ -glutamyl transferase (γ-GT).
DNA analysis. Hybridization. DNA amplification. PCR and Q-PCR. Detection strategies. Isothermic techniques for amplification of DNA. Gene expression chips.
Production of bioreactives and biomolecules. Synthesis of oligonucleotides and peptides in solid phase. Production of monoclonal and polyclonal antibodies. Synthesis of immunogenic haptens.
Immobilization of biomolecules. Strategies in solid phase in bioanalytical chemistry. Types, characteristics and nature of solid supports. Strategies for the immobilization of biomolecules in solid supports. Evaluation of nonspecific adsorption.
Labelling of biomolecules. Labelling and modifications with functional groups. Conjugation of biomolecules to different tags: enzymes, fluorophores, nanoparticles and QDs, biotin. Signal amplification techniques.
Selected videos and multimedia (password required)
– Bioanalytical Chemistry. Susan R. Mikkelsen & Eduardo Cortón. Wiley-interscience. 2004.
– Principles and Techniques of Biochemistry and Molecular Biology. 6 ª ed. Edited by Keith Wilson & John Walker. Cambridge University Press. 2006.
– ‘Bioquimica. Técnicas y Métodos’. Pilar Roca, Jordi Oliver y Ana Mª Rodríguez. Editorial Hélice. 2003.
– Principles and Practice of Bioanalysis. Edited by Richard F. Venn. Taylor & Francis, 2000
– 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.
– Skoog, Holler i Niemen. «Principios de Análisis Instrumental» 5ª edició. Ed. Mc. Graw Hill. 2001.
– Skoog, West, Holler i Crouch “Fundamentos de Química Analítica” 8ª edició. Ed. Thomsom, 2005.
– Daniel C. Harris «Análisis químico cuantitativo». 2ª ed (5ª en Ingles), Editorial Reverte 2001.
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.
– Reiner Wetermeier and Tom Naven. Proteomics in Practice: A Laboratory Manual of Proteome Analysis, Wiley-VCH Verlag-GmbH,. Weinheim, 2002.
– Rodney F. Boyer, Modern Experimental Biochemistry (Third Edition) Benjamin/Cummings, San Francisco, 2000.
– Handbook of Experimental Immunology. Vol. 1. Weir DM, ed. Oxford: Blackwell Scientific Publications, 1986.
– Rodney F. Boyer, Biochemistry Laboratory. Modern Theory and Techniques. Publisher: Benjamin Cummings. 2006.
– Jeremy W Dale and Malcom von Schantz. From Genes to Genomes: Concepts and Applications of DNA Technology. John Wiley & Sons, Ltd. 2002
– Sample Preparation Techniques in Analytical Chemistry. Somenath Mitra, ed. Wiley-Interscience: Hoboken, NJ, 2003.