TY - JOUR

T1 - Protein Quantification by Elemental Mass Spectrometry

T2 - An Experiment for Graduate Students

AU - Nehring, Andreas

AU - Tiemann, Rüdiger

AU - Schwarz, Gunnar

AU - Ickert, Stefanie

AU - Wegner, Nina

AU - Beck, Sebastian

AU - Linscheid, Michael W.

N1 - Publisher Copyright: © 2014 American Chemical Society and Division of Chemical Education, Inc. Copyright: Copyright 2014 Elsevier B.V., All rights reserved.

PY - 2014/12/9

Y1 - 2014/12/9

N2 - A multiday laboratory experiment was designed to integrate inductively coupled plasma-mass spectrometry (ICP-MS) in the context of protein quantification into an advanced practical course in analytical and environmental chemistry. Graduate students were familiar with the analytical methods employed, whereas the combination of bioanalytical assays with ICP-MS is rare. Small groups of graduate students quantified ovalbumin in hen egg white using metal-coded affinity tagging (MeCAT). Proteins were covalently labeled with lanthanide chelate complexes and quantified according to the lanthanide content by ICP-MS using internal and external standards. The results were in good agreement with reference values. As an alternative approach, a Bradford assay was used for determination of the ovalbumin content of the internal standard. The chosen workflow provides hands-on experiences for the students in principles of analytical chemistry, quantitative protein analyses, gel electrophoresis, ICP-MS, calibration, and data handling. The experiment constitutes a research-oriented approach as students apply their knowledge and skills in new contexts. (Figure Presented).

AB - A multiday laboratory experiment was designed to integrate inductively coupled plasma-mass spectrometry (ICP-MS) in the context of protein quantification into an advanced practical course in analytical and environmental chemistry. Graduate students were familiar with the analytical methods employed, whereas the combination of bioanalytical assays with ICP-MS is rare. Small groups of graduate students quantified ovalbumin in hen egg white using metal-coded affinity tagging (MeCAT). Proteins were covalently labeled with lanthanide chelate complexes and quantified according to the lanthanide content by ICP-MS using internal and external standards. The results were in good agreement with reference values. As an alternative approach, a Bradford assay was used for determination of the ovalbumin content of the internal standard. The chosen workflow provides hands-on experiences for the students in principles of analytical chemistry, quantitative protein analyses, gel electrophoresis, ICP-MS, calibration, and data handling. The experiment constitutes a research-oriented approach as students apply their knowledge and skills in new contexts. (Figure Presented).

KW - Analytical chemistry

KW - Bioanalytical chemistry

KW - Environmental chemistry

KW - Graduate education/research

KW - Hands-on learning/manipulatives

KW - Laboratory instruction

KW - Mass spectrometry

KW - Proteins/peptides

KW - Quantitative analysis

KW - Upper-division undergraduate

UR - http://www.scopus.com/inward/record.url?scp=84916608521&partnerID=8YFLogxK

U2 - 10.1021/ed500339c

DO - 10.1021/ed500339c

M3 - Article

VL - 91

SP - 2167

EP - 2170

JO - Journal of Chemical Education

JF - Journal of Chemical Education

SN - 0021-9584

IS - 12

ER -