Weighing in at 215lbs (98kg) and nearly 18ft long (5m), the female Burmese python was caught after researchers used a male "scout" to find her.
She wrestled with biologists for 20 minutes before she was "subdued", they said Wednesday.
Pythons have become pests in Florida as invasive snakes with no major predators, out-competing native species.
The serpents have been periodically found in the state since irresponsible pet owners released or allowed pet pythons to escape years ago, with many then going on to thrive in Florida's subtropical climate.
