The Digital Media Development Centre, based in Glasshouse Yard in Barbican, will house hundreds of Amazon technical staff.
They will include software development engineers, user-interface experts and graphic designers, as well as the design and development teams from Pushbutton, the TV app company, and its online entertainment service, LoveFilm.
The team will work on interactive digital services for TVs, games consoles, smartphones and PCs, as well as media projects, which it is claimed, will "benefit Amazon customers all over the world."
The centre will open within the next few months.
Paula Byrne, managing director of the centre, said that London was a "hotbed of tech talent" and testament to that was Amazon choosing it as the location for its new centre.
Byrne said: "Innovation is part of the Amazon DNA and we are creating a British centre of excellence to design and develop the next generation of TV and film services for a wide range of digital devices."
Boris Johnson, Mayor of London, said: "For this wildly successful and dynamic company to choose our city to site such an important facility is a splendid feather in our cap. We know we have the talent, the space and infrastructure to make the most of the digital economy."
The Tech City initiative was launched as a way to support and grow London's technology start-up industry, and is based around London's Shoreditch and surrounding areas, with the Old Street roundabout gaining the nickname 'The Silicon Roundabout'.
Amazon's rival Google recently set up shop in Tech City, opening a 'Campus' to support digital start-ups.Follow @shearmans