The mutual interaction of a microalga Chlorella vulgaris with four antibiotics viz. sulfamethoxazole (SMX), trimethoprim (TMP), azithromycin (AZI), and levofloxacin (LEV) individually and in mixture was studied in batch culture. SMX, TMP, and LEV stimulated algal growth, while AZI inhibited its growth. The Combination Index (CI)-isobologram indicated antagonism of the antibiotic mixture on the growth of C. vulgaris. Higher removal efficiency was observed in the mixed antibiotic than in the single antibiotic batch cultures. Biodegradation was the main antibiotic removal mechanism with a similar antibiotic biosorption pattern in single and mix antibiotic cultures. Scanning electron microscopy and Fourier transform infrared spectrophotometry showed minor biochemical alterations on algal cells surface and a stable algal population. Monod kinetics model was successfully applied to understand the growth with respect to the removal efficiency of C. vulgaris in single and mix antibiotic batch cultures. Results indicated relatively higher specific growth rate in the mix antibiotic batch culture with removal efficiency in the order of SMX > LEV > TMP > AZI. In total, 46 metabolites with 18 novel ones of the four antibiotics were identified by using high-resolution mass spectrometry based on the suspect screening approach to propose the potential transformation pathways. Most of the transformation products demonstrated lower toxicity than their respective parents. These findings implied that C. vulgaris could be an outstanding candidate for advanced treatment of antibiotic removal in wastewater.