The direct descendant of the 315 S, the 335 S made its debut at the infamous 1957 Mille Miglia with Alfonso De Portago replacing an unwell Luigi Musso at the wheel at the last minute. The car was lying third when tragedy struck near Guidizzolo, killing De Portago, his co-driver Edmond Nelson and 14 spectators. The ensuing barrage of protests brought the era of high-speed road racing to an end in Italy. The 335 S also took part in the Le Mans 24 hour, turning in the fastest lap at an average of over 120 mph. In addition to this, it took the first two places at the Venezuelan 1000 km, thus contributing to Ferrari’s victory in the Manufacturers’ World Championship.

| Type | front, longitudinal 60° V12 |
| Bore/stroke | 77 x 72 mm |
| Unitary displacement | 335.27 cc |
| Total displacement | 4023.32 cc |
| Compression ratio | 9.2 : 1 |
| Maximum power | 287 kW (390 hp) at 7400 rpm |
| Power per litre | 97 hp/l |
| Maximum torque | - |
| Valve actuation | twin overhead camshaft per bank, two valves per cylinder |
| Fuel feed | six Weber 44 DCN carburettors |
| Ignition | twin plugs, four coils |
| Lubrication | dry sump |
| Clutch | multi-plate |
| Frame | tubular steel |
| Front suspension | independent, unequal-length wishbones, coil springs, hydraulic shock absorbers |
| Rear suspension | de Dion, twin radius arms, transverse leaf spring, hydraulic shock absorbers |
| Brakes | drums |
| Transmission | 4-speed + reverse |
| Steering | worm and sector |
| Fuel tank | - |
| Front tyres | 6.00 x 16 |
| Rear tyres | 7.00 x 16 |
| Type | two-seater spider |
| Length | - |
| Width | - |
| Height | - |
| Wheelbase | 2350 mm |
| Front track | 1296 mm |
| Rear track | 1310 mm |
| Weight | 880 kg (dry) |
| Top speed | 300 km/h |
| Acceleration 0-100 km/h | - |
| 0-400 m | - |
| 0-1000 m | - |