Although tens of thousands signed the petition, most did not act upon its terms; and in August 2001, Brown and Eisen announced that they would start their own nonprofit publishing operation.

The project states (quoting the Budapest Open Access Initiative) that: "The only constraint on reproduction and distribution, and the only role for copyright in this domain, should be to give authors control over the integrity of their work and the right to be properly acknowledged and cited."

PLOS became a signatory of the SDG Publishers Compact in 2023,[9][10][11] and has taken steps to support the achievement of the Sustainable Development Goals (SDGs).

[12] In 2011, the Public Library of Science became an official financial supporting organization of Healthcare Information For All by 2015,[13] a global initiative that advocates unrestricted access to medical knowledge, sponsoring the first HIFA2015 Webinar in 2012.

[1] In 2016, PLOS confirmed that its chief executive officer, Elizabeth Marincola, would be leaving for personal and professional reasons at the end of that year.

[18] PLOS confirmed in July 2011 that it no longer relies on subsidies from foundations and is covering all of its operational costs.

[23] PLOS has its main headquarters in Suite 225 in the Koshland East Building in Levi's Plaza in San Francisco.