According to Unite the union, the hotel, an official provider to the games, does not pay staff a fair wage and commit to the agreement signed by Lord Sebastian Coe.
The agreement states that all providers of services to the Olympics should pay their workers at least the London Living Wage which is £8.30.
The protesters were from various hotels in London (not including the Holiday Inn) and were also supported by trade unionists from major cities across the world.
Diana Holland, Unite assistant general secretary, said: “The conduct of Holiday Inn in refusing to pay its staff the agreed Olympic London Living Wage makes a mockery of the ethos of the games.
“The Olympic Committee and the mayor of London must take action and hold Holiday Inn to account, and ensure that its conduct does not damage the international reputation of London as a fair world class city.”
However the hotel has said that it pays employees fairly. Laura Frith, a spokesperson for Holiday Inn, said: "To our knowledge, none of our staff are involved.
“We take part in a number of salary surveys to ensure the rates of pay for roles in London are the same as, or above, industry average.
“More than 50% of the staff working in the hotels we manage across London earn £8.31 an hour which is in line with the London Living Wage.
“We also provide all staff in the hotels we manage with meals while they are working and supply and clean their uniforms.
“We are confident that this combined pay and benefits package is equal to or above the average pay for similar roles in London.”