The largest Catholic church in the world has been disinfected ahead of the opening, with a team of suited and masked cleaners on Friday spraying every surface of the 23,000-square metre (250,000-square foot) site.
Pope Francis is not yet expected to lead any public religious ceremonies either in the basilica, which can accommodate 60,000 people, or in Saint Peter's Square, as the Vatican seeks to avoid crowds.
The Vatican State, an independent enclave in the heart of Rome, has applied the same anti-virus measures as Italy, where the official death toll from the virus stands at over 31,000.
The basilica has been closed for all religious ceremonies since March 10, when the Italian government imposed a nationwide lockdown, but individual worshippers were allowed access for private prayer.
Workers on Friday cleaned the sumptuous marble floors in the 16th century building, wielding spray canisters on the giant doors to Bernini's famous bronze Baldachin.
Vatican health official Andrea Arcangeli said the spray used was "a solution based on dilute chlorine, dosed to avoid damaging precious surfaces and works of art".
The basilica, as well as three other papal basilicas, is expected to follow a recommendation from Italy's interior ministry limiting attendance at religious celebrations in enclosed places of worship to 200 people.
Beginning Monday, the Vatican's Swiss guard will limit access to the basilica, with the help of volunteers from the Order of Malta.
The use of thermal scanners to measure the temperature of visitors is under study, but only for major religious festivals, the Vatican said.
Pope Francis will celebrate a private mass on Monday, broadcast by video, in front of the tomb of John Paul II, on the 100th anniversary of the birth of the Polish-born pontiff.
Beyond the Vatican, Catholics on Monday will be able to attend masses, baptisms, weddings and funerals in Italian churches, provided they abide by a series of measures, including wearing masks and sitting or standing well spaced apart.