The spacecraft was released from the orbiting lab at 6:40 am (1040 GMT), NASA said in a live broadcast of the event.
"Cygnus is free of the International Space Station," a NASA commentator said.
The spaceship will stay in orbit until Sunday morning, when it will fire its engines and push its way into Earth's atmosphere.
The de-orbit burn is scheduled for 8:33 am (1233 GMT) Sunday and the spacecraft should burn up at 9:11 am.
The crew on board the space station plans to document the spacecraft's plasma trail.
The resupply mission is part of a billion dollar contract with NASA for multiple journeys to the ISS.
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