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The list of Babylon 5 characters contains characters from the entire Babylon 5 universe. As the Babylon station was conceived as a political and cultural meeting place, one of the show's many themes is the cultural and social interaction between civilizations. There are five dominant civilizations represented on Babylon 5: humans, the Narn, the Centauri, the Minbari and the Vorlons; and several dozen less powerful ones. A number of the less powerful races make up the League of Non-Aligned Worlds, which assembled as a result of the Dilgar War, which occurred 30 years before the start of the series.

Jeffrey Sinclair, played by actor Michael O'Hare, is the Commander of the Babylon 5 station in season 1. After one full season, O'Hare and series executive producer/creator J. Michael Straczynski made the mutual and amicable decision for the character and actor to depart as a regular. O'Hare subsequently reprised the character of Sinclair briefly in season two and a two-episode guest appearance in season three, enabling the show to wrap up loose ends. As a result of this departure, there are several minor inconsistencies between the first season and the remainder of the show, most visibly in "And the Sky Full of Stars" and "Babylon Squared". At O'Hare's personal request, the full reasons for his departure from the show were kept secret until after his death in 2012. The following year, Straczynski revealed that O'Hare struggled with delusion and paranoia due to mental illness, which ultimately prevented him from continuing to act. However, Straczynski emphasized that O'Hare's fans, particularly those of his role as Sinclair, had helped him cope with his struggle in ways medication never could.

The character's backstory is given as having been born on Mars Colony. In an early episode of season one, Sinclair states that his family had been pilots "ever since the Battle of Britain" and Sinclair's father was a fighter pilot for EarthForce who participated in the Battle of Balos, the last engagement of the Dilgar Invasion. Sinclair continued the tradition and became a fighter pilot. Sinclair enlisted in EarthForce in 2237. During his time at EarthForce Academy, he met Catherine Sakai, with whom he had a relationship. After a year of living together, the two of them broke up, but continued to see each other off and on through 2258, when they became engaged. Sakai went missing in late 2259, while on a mission for the Rangers. The season one episode "By Any Means Necessary" establishes that Sinclair received Jesuit education as a young man.

In 2240, Sinclair was promoted to fighter pilot, continuing a Sinclair family tradition that had extended as far back as the Battle of Britain. Less than a year later, Sinclair was promoted to squadron leader; due to his rapid rise through the ranks, the rumor of the day was that Sinclair was on the fast track to making Admiral. As squadron leader, Sinclair fought at the Battle of the Line, the last major battle in the Earth-Minbari War. During the course of the battle, his squadron was destroyed by the Minbari, and his fighter was badly damaged. In a last act of defiance, Sinclair attempted to ram one of the Minbari cruisers. He failed in this when he was captured by another Minbari cruiser for interrogation by the Grey Council. The Council's Triluminary detected Valen's DNA in Sinclair, so they assumed, to their profound shock, that he possessed the soul of Valen, a hero of the Minbari who led them to victory 1,000 years ago against the Shadows. It was concluded by the Grey Council that Minbari souls were being born into human bodies. The discovery of what Sinclair possessed led the Minbari to surrender and return Sinclair to his fighter, the memory of his time aboard the Minbari cruiser blocked (though this block would not be permanent and would break down years later). Sinclair — and the Earth Alliance — believed that he had blacked out from the acceleration. When Babylon 5 was brought into operation in 2257, Sinclair was selected by the Minbari to command the newly constructed station. He was selected over many more senior officers, including Colonel Ari Ben Zayn, all of whom had been vetoed by the Minbari (they had stipulated that they should approve the choice of Station Commander, as they had shared the cost of construction).

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