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NIST Spectroscopic Measurement Standards



Paul C. DeRose, Kenneth D. Cole, Aaron Urbas, Steven J. Choquette, Evelyn Solis, Erica V. Stein, John E. Schiel, Brian Lang, Hua-Jun He


Ultraviolet (UV) absorbance measurements provide a rapid and reliable method to determine protein concentrations. The National Institute of Standards and Technology (NIST) has developed Standard Reference Material (SRM) 2082as a pathlength standard for UV absorbance measurements for use with the new generation of microvolume spectrophotometers and short-pathlength cuvettes. Short pathlengths are used with high-concentration targets to ensure that absorbance values are within the optimal range. The short-pathlength instruments and cuvettes also reduce the required volumes to conserve valuable samples. The authors compared the results obtained with high-quality dual-beam spectrophotometers and short-pathlength cuvettes to the results obtained from a microvolume spectrophotometer and a microvolume plate reader. SRM 2082 can be used to accurately calculate pathlength values, thereby increasing the accuracy in subsequent measurements using the short-pathlength cuvettes and microvolume absorbance instruments. RM 8671 (reference material, the NISTmAb) can then be used to ensure the accuracy and reproducibility of protein concentration measurements by providing an industrially relevant reference material, a well-characterized monoclonal antibody.
BioPharm International


absorbance, standard, monoclonal antibody, pathlength, protein, measurement, biopharmseutical, reference material


DeRose, P. , Cole, K. , Urbas, A. , Choquette, S. , Solis, E. , Stein, E. , Schiel, J. , Lang, B. and He, H. (2018), NIST Spectroscopic Measurement Standards, BioPharm International (Accessed April 15, 2024)
Created May 1, 2018, Updated October 13, 2022