The sold-out game against the Utah Jazz starts at 7:30 p.m. and will be the most heavily covered regular season game in NBA history. More than double the amount of press credentials were given out to media members from all over the world.
Tickets for the game are priced for as much as $25,000 on Stub Hub, which is the most expensive regular season ticket average for any sport ever.
Including playoff games, this will be Bryant's 1,556th game of his career, each one donning a Lakers uniform. The five-time champion will cap his legacy with a 25.0 scoring average - regardless if he scores any points during Wednesday's game.
The first three free throws he shoots at his last game will be the 10,000th of his career, which is something only four other players have ever done.
In a letter posted online, Magic Johnson issued a heartwarming tribute to Bryant.
"Every night you played, I couldn't wait to watch knowing that I would witness an unbelievable shot or move that I had never seen on the court," the letter said. "I marveled at the skills and loved the energy you brought to the game wearing that purple and gold uniform. You never disappointed the city, Laker fans..."
In addition to avid fans attending the game, Bryant's former teammates Shaquille O'Neal and Rick Fox will be in the crowd, as well as Red Hot Chili Peppers bassist Flea among other celebrities.
In honor of Bryant's retirement, the Pico-Union Metro Station will be dubbed the "Kobe Station" for the day as people head out to his last game.
In preparation for the massive event, the Staples Center set up hundreds of limited edition items that went on sale at 8:24 a.m. - an homage to his Lakers jersey numbers.
Items included a lambskin jacket marked with Bryant's No. 24, T-shirts, hats and jerseys.
Bryant announced his retirement last November in a poem titled "Dear Basketball," which was published on the website The Player's Tribute.
A video tribute to his illustrious career will be shown before player introductions during Wednesday's event. Video tributes to him will also be played out during timeouts over the course of the game.
When it concludes, Bryant will speak with the crowd. All fans in attendance will receive a commemorative program, T-shirt and ticket lanyard.
City News Service and the Associated Press contributed to this report.