Enzymes have essential roles in catalyzing biological reactions and maintaining metabolic systems. Many in vitro enzymatic bioassays have been developed for use in industrial and research fields, such as cell biology, enzyme engineering, drug screening, and biofuel production. Of note, many of these require the use of high-throughput platforms. Although the microtiter plate remains the standard for high-throughput enzymatic bioassays, microfluidic arrays and droplet microfluidics represent emerging methods. Each has seen significant advances and offers distinct advantages; however, drawbacks in key performance metrics, including reagent consumption, reaction manipulation, reaction recovery, real-time measurement, concentration gradient range, and multiplexity, remain. Herein, we compare recent high-throughput platforms using the aforementioned metrics as criteria and provide insights into remaining challenges and future research trends. Advances in microtiter plates, microfluidics arrays, and droplet microfluidics show unique advantages for high-throughput enzymatic bioassays regarding key performance metrics, including reagent consumption, reaction manipulation, reaction recovery, real-time measurement, concentration gradient range, and multiplexity. Advances in microtiter plates focus on their integration with accessory systems to improve their overall performance, such as sensitivity and reaction rates. Advances in microfluidic arrays aim to reduce reagent consumption, increase assay throughput, and enable multi-step bioassays and reaction recovery. Advances in droplet microfluidics enable binary detection with integrated sorting, kinetic screening under continuous flow, real-time measurement via in situ droplet trapping, and multiplexing and combinatorial screening. [ABSTRACT FROM AUTHOR]