Downstream processing of antibiotics is usually conducted through conventional solvent extraction. Several limitations of the conventional solvent extraction are formation of stable emulsion, solvent loss using volatile solvents, low extraction yield, and unable to separate amphoteric antibiotics which are barely soluble in organic solvents. Reverse micelle extraction is an attractive alternative for replacing conventional solvent extraction of antibiotics. It has several advantages: high selectivity, high mass and activity recovery, simple operation, cost effective, easy to scale up, using non-toxic substances, and can design to be as environmental friendly as possible by using biosurfactants. This paper briefly introduces reverse micelle extraction and reviews several crucial factors that should be manipulated during operation to achieve optimum recovery. The process can be easily optimized by adjusting those factors.