Chemotherapy constitutes the main strategy in management of breast cancer (BC). Lack of specificity and high burden of adverse effects of chemotherapeutic agents remain the most important impediments to successful treatment of BC patients. Folate receptor α (FRα) could be very promising for therapeutic targeting in this type of cancer. In this study, ß-lactoglobulin nanoparticles (BNPs) conjugated with folic acid and loaded with doxorubicin (FDBNPs) were prepared. Various characterization techniques were applied to determine the size, polydispersity and doxorubicin loading of prepared FDBNPs in comparison with doxorubicin-loaded BNPs (DBNPs). The results showed that FDBNPs are 109.77 ± 2.80 nm in diameter with well dispersed and spherical shapes. The biodegradation of FDBNPs in the presence of trypsin enzyme and in PBS at different pH (4 and 7) was spectrophotometrically monitored and the results showed that the FDBNPs with encapsulation efficiency of 68.82%±1.76% could deliver doxorubicin at clinically relevant doses. Effects of DBNPs and FDBNPs against MCF-7 and MDA-MB-231, BC and triple negative BC (TNBC) cell lines, respectively, showed significant inhibition of cell proliferation as well as induction of apoptosis. Based on these findings, FDBNPs with facilitated drug release and targeted doxorubicin delivery capacities could have high therapeutic potential for BC and TNBC.