Application of agro-industrial by-products for the production of lactic acid was studied in this paper. Brewer's spent grain (BSG), malt rootlets (MR), brewer's yeast (BY), and soy lecithin (SL) were used as raw materials in l-(+)-LA fermentation by free and immobilized Lactobacillus rhamnosus ATCC 7469. The BSG, solid remains after BSG and MR hydrolysis (BSGMRSR), and MR were evaluated as carriers for batch and repeated batch fermentations with immobilized cells. During batch fermentations with immobilized cells, high cell viability (10 to 11logCFU/g) was achieved on all carriers. In batch fermentation with BSG as a carrier, the highest LA yield of 93.79% and volumetric productivity of 1.15g/L/h were obtained. Furthermore, very high LA yield (95.46%), volumetric productivity (1.98g/L/h) and L. rhamnosus viability (11.5logCFU/g) were achieved in repeated batch fermentations with the cells immobilized on this carrier. The immobilized cells showed high survival rate (94-95%) during exposure to simulated gut condition. Based on the analysis of BSGMRSR, and BY solid remains, and on in vitro evaluation of the probiotic characteristics of immobilized cells, it was observed that they could satisfy the recommendations for high-quality feed preparation.