An Excel fitting routine for correcting protein absorption spectra for scatter

被引：2

作者：

Pilkington, Tyson J. ^{[1
]}

Bignucolo, Matthew W. ^{[1
]}

Ainsworth, Jack P. ^{[1
]}

Siemann, Stefan ^{[1
,2
]}

机构：

[1] Laurentian Univ, Sch Nat Sci, Sudbury, ON, Canada

[2] Laurentian Univ, Sch Nat Sci, 935 Ramsey Lake Rd, Sudbury, ON P3E 2C6, Canada

来源：

ANALYTICAL BIOCHEMISTRY | 2023年 / 678卷

基金：

加拿大自然科学与工程研究理事会;

关键词：

Protein absorption; UV-Vis spectroscopy; Rayleigh scatter; Mie scatter; Scatter correction; ULTRAVIOLET-ABSORPTION; LIGHT-SCATTERING; QUANTITATION; ASSAY;

D O I：

10.1016/j.ab.2023.115269

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Protein concentrations are often determined in a non-destructive manner by measuring the absorbance at 280 nm. However, light scattering in protein samples can complicate such assessment. We here describe a simple Excel Solver-based fitting routine to correct full protein UV absorption spectra for both Rayleigh and Mie scattering. Using samples displaying various degrees of natural and artificially induced scattering, we show that our multi-wavelength fitting method is not only capable of aiding in the determination of protein concentrations but can also be employed in the spectral analysis of protein structural changes that are accompanied by alter-ations in scatter intensity.

引用

页数：4

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