Casein is the major protein fraction in milk, and its cross-linking has been a topic of scientific interest for many years. Enzymatic cross-linking has huge potential to modify relevant techno-functional properties of casein, whereas non-enzymatic cross-linking occurs naturally during the storage and processing of milk and dairy products. Two size separation techniques were applied for characterisation of these reactions: gel electrophoresis and size exclusion chromatography. This review summarises their separation principles and discusses the outcome of studies on cross-linked casein from the last ~20 years. Both methods, however, show limitations concerning separation range and are applied mainly under denaturing and reducing conditions. In contrast, field flow fractionation has a broad separation range and can be easily applied under native conditions. Although this method has become a powerful tool in polymer and nanoparticle analysis and was used in few studies on casein micelles, it has not yet been applied to investigate cross-linked casein. Finally, the principles and requirements for absolute molar mass determination are reviewed, which will be of increased interest in the future since suitable calibration substances for casein polymers are scarce.

In situ produced extracellular polysaccharides (EPS) from lactic acid bacteria are generally known to affect the texture of fermented dairy products; however, the interplay between EPS and product properties is still poorly understood. The aim of this study was to establish a relationship between concentration and properties of EPS, and gel formation of milk analysed by noninvasive Multispeckle Diffusing Wave Spectroscopy. Twenty Streptococcus thermophilus strains were classified with respect to EPS concentration (8–126 mg GE/kg) and ropiness (thread length: 15–80 mm). Five groups identified by cluster analysis demonstrate the high strain-to-strain variability even within one species of lactic acid bacteria. Results from acidification and gelation experiments averaged per cluster indicate that fermentation time and gel stiffness is higher for strains that produce ropy EPS. A further increase in gel stiffness was detected for strains that also produced cell-bound EPS, which underlines the importance of both ropy and cell-bound EPS for improving acid gel properties. The results may be helpful for a proper selection of EPS-producing starter cultures.