Google has lodged its appeal to the EU's Luxembourg-based European Court of Justice over the €2.4 billion fine imposed on it by the European Commission Competition Directorate for favouring its shopping service in search results.
Google is now set for a legal showdown in Brussels that could take years to resolve, and add further strain on already tense relations between the company and the EU.
Following the ruling, Google said it "respectfully" disagreed and was considering whether to appeal.
Google has already submitted a rough draft to regulators over changes it must make by September 28 to avoid further fines, which should include giving equal treatment to rivals.
Brussels had accused Google of giving more preference to its own services in the search results to the determinant of other price comparison sites, such as TripAdvisor and Expedia.
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However, setting up a court battle between Brussels and the internet giant could take years to resolve and make already tense relations between Europe and the U.S. tech giant even more fraught, AFP reported.
Margrethe Vestager, the EU's competition commissioner had said that Google's activity was "illegal under European Union antitrust rules". "And most importantly, it denied European consumers a genuine choice of services and the full benefits of innovation".
The EU Court of Justice told a lower tribunal last Wednesday to re-examine U.S. chipmaker Intel's appeal against a 1.06 billion euro fine, dealing a rare setback to the Commission.
The EU is also investigating whether Google tried to squeeze out its rivals in online search advertising and through its Android mobile operating system.