In this study, four microalgal strains were evaluated for their biomass production capacity and macromolecule biosynthesis. These include three strains from the phylum Chlorophyta: Monoraphidium sp. LB2PC 0120, Stichococcus sp. LB2PC 0117, and Tetraselmis sp. LB2PC 0320, and one strain from the phylum Haptophyta: Isochrysis sp. LB2PC 0220. The experiments were conducted under typical laboratory-scale setups. Additionally, phylogenetic analysis based on the 18-28 S rRNA internal transcribed spacer (ITS) was performed to validate the taxonomic identity of the strains. Each strain was exposed to four different cultivation conditions based on two levels of illumination intensity [25-(LI) and 50-(HI) µmol m s] and nitrogen loading [100-(LΝ) and 300-(HΝ) mg NaNO L] in a full factorial design. All the microalgae achieved maximum biomass production under HI-HN conditions, which amounted to 1495, 919, 844, and 708 mg/L for Monoraphidium sp. LB2PC 0120, Stichococcus sp. LB2PC 0117, Tetraselmis sp. LB2PC 0320 and Isochrysis sp. LB2PC 0220, respectively, after 16 days of cultivation. Among them, Stichococcus sp. LB2PC 0117 had the highest protein content (49.9% wt.) under LI-HN conditions and Monoraphidium sp. LB2PC 0120 had the highest lipid content (44.3% wt.) under HI-LN conditions. Both Monoraphidium sp. LB2PC 0120 and Tetraselmis sp. LB2PC 0320 accumulated the highest carbohydrate content (~ 37% wt.) under LI-LN and HI-LN conditions, respectively. Based on biomass and macromolecule production, Monoraphidium sp. LB2PC 0120 was identified as the most promising candidate for upscaling studies, expecting its highly manipulatable compositional profile to support multiple applications in the food industry, rendering this microalga a valuable resource.