Development of a univariate membrane-based mid-infrared method for protein quantitation and total lipid content analysis of biological samples.
Biological samples present a range of complexities from homogeneous purified protein to multicomponent mixtures. Accurate qualification of such samples is paramount to downstream applications. We describe the development of an MIR spectroscopy-based analytical method offering simultaneous protein quantitation (0.25-5 mg/mL) and analysis of total lipid or detergent species, as well as the identification of other biomolecules present in biological samples. The method utilizes a hydrophilic PTFE membrane engineered for presentation of aqueous samples in a dried format compatible with fast infrared analysis. Unlike classical quantification techniques, the reported method is amino acid sequence independent and thus applicable to complex samples of unknown composition. By comparison to existing platforms, this MIR-based method enables direct quantification using minimal sample volume (2 µL); it is well-suited where repeat access and limited sample size are critical parameters. Further, accurate results can be derived without specialized training or knowledge of IR spectroscopy. Overall, the simplified application and analysis system provides a more cost-effective alternative to high-throughput IR systems for research laboratories with minimal throughput demands. In summary, the MIR-based system provides a viable alternative to current protein quantitation methods; it also uniquely offers simultaneous qualification of other components, notably lipids and detergents.
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Ivona
0000-0003-0010-7986
I
Strug
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Christopher
0000-0002-3760-4735
C
Utzat
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Amedeo
A
Cappione
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Sara
0000-0002-8343-4909
S
Gutierrez
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Ryan
0000-0002-0052-0554
R
Amara
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Joseph
J
Lento
Institute for Organic Chemistry and Biochemistry, Technical University Darmstadt, 64289 Darmstadt, Germany.
Florian
0000-0003-4082-2386
F
Capito
Merck KGaA, Frankfurter Straße 250, 64293 Darmstadt, Germany.
Romas
0000-0002-3567-8765
R
Skudas
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Elena
E
Chernokalskaya
EMD Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923, USA.
Timothy
0000-0002-3898-1743
T
Nadler