AbstractDifferent immobilization methods were adapted to the 96‐well microtiter plate scale using esterases as model enzymes. The methods tested were based on adsorption, coprecipitation, aggregation and covalent bonding. The protein covered microcrystals proved to be the best method in terms of yield and expressed activity for the test reaction, which was the alcoholysis of p‐nitrophenyl acetate with 1‐propanol under anhydrous conditions.