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Development of an improved in vitro protein digestibility (IVPD) model system

Educational level:Master thesis
Target group:MSc Food Innovation and Health
MSc Food Science and Technology
Main area:Ingredients and Dairy Technology
Chemometrics and Analytical Technology
Project description:

Protein digestibility is one of the most important aspects of nutritional quality, since it largely determines the bioavailability of proteins in food. The “gold standard” for assessing protein digestibility relies on physiological studies on human subjects. However, in vitro protein digestibility (IVPD) models may serve as an important supplement to in vivo studies due to their simplicity, rapidness, and cost-efficiency. IVPD model systems attempt to simulate the gastro-intestinal digestion of humans upon food consumption. Typically, static IVPD model systems are employed under fixed experimental conditions such as pH, temperature, hydrolysis time, and enzyme:substrate ratios (E:S ratios). Static IVPD model systems essentially consist of two parts: (1) protein digestion by enzymatic hydrolysis, followed by (2) quantification of the degree of protein digestion using various detection methods such as TNBS, OPA, and pH-stat. All these methods have certain strengths and weaknesses in relation to their application potential.

In our laboratory, we have developed a static IVPD model system simulating the gastro-intestinal digestion of human adults. This model system involves enzymatic protein hydrolysis and quantification using a TNBS-based assay. It is widely used, well-functioning, and has been published in several scientific papers. Now we would like to further improve the method, perhaps also redesign some of the procedural steps and supplement it with additional add-on analyses. Therefore, the aim of this MSc project is mainly to develop an improved adult IVPD model system based on best practices within the currently used method. The practical work will essentially include method development, optimization, and validation, as well as data handling and statistical analysis. Multiple parameters will be investigated such as intra-day and inter-day variation of the method under different assay conditions. Experimental conditions such as E:S ratios, enzyme combinations, and hydrolysis times will altered to gain an optimized IVPD model system. Detection methods used for quantification of protein digestibility will be compared, fx between the TNBS and OPA method. Supplementary add-on analyses for studying the extent of protein digestion will be explored such as SDS-PAGE and LC-MS. Lastly, trials will be commenced to develop an infant model system based on adjustment of the adult IVPD method.

Keywords:Nutritional quality, Protein digestion, In vitro protein digestibility (IVPD), Gastro-intestinal digestion, Analytical method development, optimization, and validation
Supervisor(s): Marcel Skejovic Joehnke, René Lametsch
Last edited:28-11-2019