The petition, to stop the East Coast line being re-privatised, is being handed today to shadow rail minister Lilian Greenwood and Green Party MP Caroline Lucas.
The line has been run in the public sector since autumn 2009 but is due to go back into private ownership in early 2015.
The petition has been signed by more than 23,000 rail passengers. Also, more than 60 MPs from Labour, the Liberal Democrats, Green and Scottish National (SNP) parties have signed an early day motion calling on the Government not to re-privatise the line.
The campaign is supported by RMT union general secretary Bob Crow and TSSA union general secretary Manuel Cortes. They argue that a publicly-run East Coast is better value for money than a line operated in the private sector.
Campaigners have highlighted figures announced last week which showed that East Coast paid £208 million in premium and dividend payments to the Treasury in the last financial year (2012/13).
The campaigners also point out that figures from the Office of Rail Regulation show that East Coast received the lowest public subsidy of any rail operator by some considerable margin.
In addition, those keen to keep East Coast publicly run say that the Government is rushing through this particular re-franchising so it takes place before the 2015 general election.
Ms Greenwood said: "East Coast has gone from strength to strength since 2009. Passenger satisfaction is at record levels, all profits have been reinvested in the service and £600 million has been returned to the taxpayer already.
"It makes no sense to waste public money and government time on selling off East Coast instead of getting to grips with the cost of living crisis. David Cameron should now take note of this petition and cancel this ideologically-driven privatisation."
Action for Rail spokesperson and TUC general secretary Frances O'Grady said: "Privatising the East Coast Main Line defies all logic. Since it was re-nationalised the line has made a large contribution to the taxpayer, bringing in over £600m already.
"By taking the East Coast out of public ownership the Government will be passing the income the line raises into the pockets of corporate shareholders, when it should be using the cash to reduce rail fares and improve services."
A Department for Transport spokesman said: "The independent Brown Review into rail franchising was clear that franchising remains the best way to secure services for passengers.
"A strong private-sector partner will not only provide certainty of ownership for the East Coast franchise but will be best placed to build on the significant investment planned for the route, delivering benefits to passengers and taxpayers for years to come."