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The link between the O2 Arena in Greenwich and the ExCel exhibition centre at the Royal Docks will be capable of carrying up to 2,500 people an hour.
A single adult fare on the pay-as-you-go Oyster card will cost £3.20 while the cash fare will be £4.30, Transport for London (TfL) said.
Passengers will also be able to make a non-stop round trip on the cable car, with views of the City, Canary Wharf, the Thames Barrier and the Olympic Park, at a cost of £6.40 with Oyster.
The service will operate through the week from 7am until 9pm.
Dubai-based airline Emirates is sponsoring the cable car for 10 years at a cost of £36 million.
The total cost of the project is about £60 million.