NSTI BioNano 2010

Simplified Quantitation of Proteins at A280 using a Low Cost, One Millimeter Matched Pathlength Plastic Cuvette

L.M. Levine, J. McDowell, N.T. Wong, J. Beebe, J. Horne, P. Schultz
ALine, Inc., US

Keywords: matched pathlength, short pathlength cuvette, A280 protein absorbance, protein quantitation, plastic, UV transparent

Abstract:

Routine measurement of protein concentration in a production environment is typically done using a 1 cm pathlength quartz cuvette. The protein to be measured requires dilution to be within the dynamic range of the spectrometer. Dilution involves an additional step in the concentration measurement process, and often leads to random or systematic errors that skew the measurement and either under or over estimate the concentration. These errors in concentration can increase manufacturing variability for diagnostic tests that require a known quantity of protein to be incorporated into a test device. To avoid experimental error due to dilution, 1 mm pathlength quartz cuvettes have been used, however the extra steps required for cleaning can also introduce errors into the measurement. The technician might not clean the contents completely, or be unable to clean it due to non-reversible adsorption of proteins on the quartz surface. To avoid these issues, a single-use 1 mm pathlength cuvette has been developed and shown to permit quantitation of protein concentrations with accuracy and precision similar to that of a 1 mm quartz cuvette.
 
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