The aim of this study was to determine the effect of addition of alfalfa protein concentrate (APC) at the dose of 15 g or 30 g per 1 kg of complete feed ration for the native Polbar breed on selected production traits and the fatty acid profile in the yolk of raw, hard-boiled or freeze-dried eggs. Laying hens were assigned randomly to 3 experimental groups, each comprising 30 birds. The control group received a standard diet without the APC addition and the experimental groups received APC, which partially replaced postextraction soybean meal. Egg laying performance, feed intake, and feed conversion ratio were controlled throughout the experiment. At 33 wk of age, 45 eggs were randomly selected for assessment of the quality of the egg content and eggshell, and 30 eggs were taken for each of the cooking and freeze-drying treatments. The fatty acid composition was determined in the yolks of hard-boiled, freeze-dried, and raw eggs. There was no effect of the APC addition on the laying performance, feed intake and feed conversion ratio, and a majority of egg quality traits. Hens fed with the APC laid eggs with a darker colored eggshell and yolk and a slightly lower breaking strength. The yolks in the eggs from hens receiving the APC addition were characterized by distinctly higher content of polyunsaturated fatty acids (PUFA). The group fed with a higher dose of APC produced eggs with a substantially lower level of saturated fatty acids (SFA). Boiling resulted in an increase in the SFA content and a decline in the level of PUFAs and carotenoids. Freeze-drying led to an increase in the total SFA content and a decrease in the level of n-3 PUFA. The APC addition to feed can replace the genetically modified soybean meal without reducing the values of production traits and egg quality and with a beneficial effect on the yolk color and fatty acid profile.