Antibiotic residues in milk may pose a risk for public health and for the dairy industry, especially in fermented products such as cheeses. The objective of this study was to analyze the effect of β-lactams (amoxicillin, benzylpenicillin, and cloxacillin) in goat’s milk on the biochemical and texture characteristics of semi-hard Tronchón cheese. Milk from Murciano-Granadina goats with a good health status and not having received any veterinary drugs was used. The milk was spiked with β-lactam concentrations equivalent to their maximum residue limit (MRL) and ripened cheeses were made. One batch of cheeses from raw antibiotic-free milk was used as control. The proteolysis level was determined by free amino acids content (FAA, mg of leucine/g of cheese), the lipolysis level measured as the free fatty acids concentration (FFA, mEq KOH/100 g of fat) and texture parameters assessed by texture profile analysis (TPA: hardnesss, adhesiveness, springiness, cohesiveness, chewiness). Also, the residual concentration of antibiotics by HPLC-MS/MS in cheeses ripened for different periods (1, 30, and 60 d) was studied. The presence of amoxicillin in milk at MRL affected the FFA concentration in cheeses, being lower in antibiotic-spiked cheeses compared with control cheeses (2.21 vs. 2.67 mEq KOH/100 g of fat). However, the presence of benzylpenicillin in milk did not have any negative effects on the cheese properties. While, in cheeses made from cloxacillin spiked milk FFA concentration being lower than in control cheeses (2.40 vs. 2.67 mEq KOH/100 g of fat), also springiness was affected likewise (0.60 vs. 0.63). Amoxicillin (<2 µg/kg), benzylpenicillin (4.8 ± 1.8 µg/kg) and cloxacillin (28.8 ± 1.7 µg/kg) residual amounts were quantified at beginning of ripening (1 d), while antibiotic residues were not detected in the cheese at the end of ripening (60 d). It can be concluded that the β-lactam residue concentrations in matured goat’s cheeses do not represent a potential public health risk. Nevertheless, similar studies with other antibiotic groups and using different cheese-making processes are recommended.