"There is no justification for the closure of the Rafah crossing," Hamad al-Raqb, a spokesman for the protesters, said in a speech marking the beginning of a four-day sit-in.
"It is unbecoming of the historic relations between Egyptians and Palestinians," he insisted.
Leading Hamas figures and MPs attended the launch ceremony.
Egyptian authorities have tightened their control over the border with the Hamas-run Gaza Strip since last July's ouster of elected president Mohamed Morsi by the Egyptian army.
In recent weeks, Egypt has repeatedly closed the Rafah crossing, which – due to an ongoing, eight-year Israeli siege – represents Gaza's only window to the outside world.
The Egyptian army has also launched a campaign aimed at destroying the network of tunnels linking Gaza to Egypt's Sinai Peninsula, which in recent years had been used to bring vital commodities into the besieged coastal enclave, home to nearly 2 million Palestinians.
Hamas, an ideological offshoot of the Muslim Brotherhood, the group which propelled Morsi to power in the 2012 elections, has controlled the Gaza Strip, since 2007.
Deadly clashes are the latest in months of unrest triggered by President Pierre Nkurunziza's successful bid to win a third term in office
'I think the reputation of German industry, the confidence in the German economy, is not so shaken that we are no longer considered a good place to do business,' Merkel says
Israeli police says attacker shot after stabbing Israelis in Jerusalem's old city
3 people have been arrested in Mogadishu after police raided a private television channel.
A new exhibition at the Northern Illinois University highlights the material culture of indigenous Muslims in the Philippines.
The German cabinet is keeping minimal staff at it Uzbekistan air base, paying $35 million a year.
Reported shooting comes as Israeli forces search for killers of settler couple
Palestinians were protesting against the raid in the territory's main northern city of Nablus hurled stones at soldiers, who responded with rubber bullets, tear gas and live ammunition, the sources said.
UN secretary general has appealed for an end to fighting as experts fear looming humanitarian disaster with Libya factions missing the ceasefire deadline.
Bomb blasts have ripped through Abuja killing 15 people
Seven opposition candidates call for delaying Oct. 11 presidential polls in light of ‘electoral irregularities’
The banning of Tajikistan's main opposition party has drawn condemnation from the United Nations.
8 ministries, 30 departments have been cut in a shakeup that will slash ministers' pay
Belgian families open doors to refugees in Brussels to save them from the cold as they process asylum applications
Island solution not to create losers and winners, but 'winners on both sides', says Turkish Cypriot leader, Akinci, while the Greek Cypriot leader agreed saying that a window for a deal exists