It happened in the Fort Erie-bound lanes of the Queen Elizabeth Way, near Fruitland Road, at around 4:40 a.m.
According to police, a vehicle travelling in the wrong direction collided with another vehicle.
The driver of the vehicle travelling in the correct direction was killed in the crash. He has since been identified by provincial police as 38-year-old Khoi Dinh Le, of Cambridge.
The driver of the vehicle travelling in the wrong direction has been charged in connection with the crash.
Ancaster resident Vukasin Krsmanovic faces charges including impaired operation of a motor vehicle causing death, over 80 mg causing death, and dangerous driving causing death.
He is scheduled to appear in court in Hamilton on April 15.
Anyone with information is asked to contact police at 905-681-2511.