Fluorescence-based soft-sensor for monitoring beta-lactoglobulin and alpha-lactalbumin solubility during thermal aggregation.
A soft-sensor for monitoring solubility of native-like alpha-lactalbumin (alpha-LA) and beta-lactoglobulin (beta-LG) and their aggregation behavior following heat treatment of mixtures under different treatment conditions was developed using fluorescence spectroscopy data regressed with a multivariate Partial Least Squares (PLS) regression algorithm. PLS regression was used to correlate the concentrations of alpha-LA and beta-LG to the fluorescence spectra obtained for their mixtures. Data for the calibration and validation of the soft sensor was derived from fluorescence spectra. The process of thermal induced aggregation of beta-LG and alpha-LA protein in mixtures, which involves the disappearance of native-like proteins, was studied under various treatment conditions including different temperatures, pH, total initial protein concentration and proportions of alpha-LA and beta-LG. It was demonstrated that the multivariate regression models used could effectively deconvolute multi-wavelength fluorescence spectra collected under a variety of process conditions and provide a fairly accurate quantification of respective native-like proteins despite the significant overlapping between their emission profiles. It was also demonstrated that a PLS model can be used as a black-box prediction tool for estimating protein aggregation when combined with simple mass balances.
Department of Chemical Engineering, University of Waterloo, Waterloo, ON, Canada, N2L 3G1.
Rand
R
Elshereef
Hector
H
Budman
Christine
C
Moresoli
Raymond L
RL
Legge
D000465:Algorithms; D003460:Crystallization / Q000379:methods; D006358:Hot Temperature; D007768:Lactalbumin / Q000032:analysis* / Q000737:chemistry*; D007782:Lactoglobulins / Q000032:analysis* / Q000737:chemistry*; D046912:Multiprotein Complexes / Q000032:analysis; D011485:Protein Binding; D012995:Solubility; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D014159:Transducers
D007782:Lactoglobulins; D046912:Multiprotein Complexes; D007768:Lactalbumin