Angry Birds: Black bird (choleric), white bird (melancholic), red bird (phlegmatic), and yellow bird (sanguine).
The four spirits of the Book of Prophecy in Avalon Code can be identified as this: Rempo (choleric), Ur (melancholic), Neaki (phlegmatic), and Mieli (sanguine).
In Blaze Union, the first five party members: Garlot (choleric), Jenon (melancholic), Medoute (phlegmatic), Siskier (sanguine), and Aegina (leukine). And after the party dynamics suffer an upheaval, things shake down like so: Gulcasa and Leon (choleric), Nessiah (melancholic), Baldus (phlegmatic), Emilia (sanguine), and Aegina is... still leukine.
Borderlands: Mordecai (melancholic), Lilith (phlegmatic), Roland (choleric), and Brick (sanguine).
Borderlands 2: Axton (sanguine with a bit of choleric), Maya (melancholic), Salvador (choleric), Zer0 (phlegmatic). With DLC, there's also Gaige and Krieg, who are Sanguine and Choleric respectively.
Hyperion: Handsome Jack (choleric), Mr. Blake (melancholic), Professor Nakayama (sanguine), Angel (phlegmatic).
Catherine, in terms of Vincent and his three bros that he hangs out with at the Stray Sheep. Orlando (choleric), Jonny (melancholic), Vincent (phlegmatic), and Toby (sanguine).
Melancholic: Hinata, Koizumi, Pekoyama and Gundam.
Phlegmatic: Komaeda, Nanami and Sonia.
The Love Interests in Dragon Age II: Isabela (sanguine), Merrill (choleric), Anders (melancholic), Fenris (phlegmatic with a dash of melancholic).
The four playable characters of Drakengard, though at times they muddle about with their humors, tend to fall into distinct ones: Arioch (sanguine), Caim (choleric), Leonard (melancholic), and Seere (phlegmatic).
Due to the author being interested with the subject, MARDEK RPG does this. The author believes everyone has a primary and a secondary temperament, though. Perhaps the clearest ingame example is Legion: a construct created by an inventor. It was animated with dark magic, with the inventor saying the incantation - due to his stuttering, though, four souls entered the body instead of one - one for each temperament.
Mass Effect 2 has the starting four. Shepard (choleric), Miranda (melancholic), Jacob (phlegmatic), and Joker (sanguine).
Mass Effect 1 and Mass Effect 3 squads are often comprised this way: Shepard and Javik(choleric), Garrus and Liara (melancholic) and Tali and EDI (phlegmatic). Likewise, James and Ashley (sanguine) and Kaiden (neutral phlegmatic).
Mega Man (Classic) series: Forte/Bass (choleric/sanguine), Proto Man/Blues (melancholic/choleric), Mega/Rock Man (melancholic/sanguine), and Duo (phlegmatic/leukine).
Mega Man Battle Network: Dex and his Navi GutsMan.EXE (choleric), Chaud and Glyde.EXE (melancholic), Mayl and MegaMan.EXE (phlegmatic), Yai and her Navi Roll.EXE (sanguine), and Lan (leukine).
Mega Man X Maverick Hunters: The fast-action Anti-Hero Zero (choleric), the conscientious defender X (melancholic), the calm and fatherly Signas (leukine/phlegmatic), and the flamboyant Axl (sanguine).
The villains: Vile is choleric (and dang!!); Lumine is melancholic; Dynamo is sanguine; and Sigma is leukine.
Mega Man X2's X-Hunters, plus Vile as the solo X-Hunter in his own right: Vile (choleric/melancholic), Agile (melancholic/sanguine), Violen (sanguine/choleric), and Serges (phlegmatic/leukine).
Capcom never made clear whether classic Mega/Rock Man is the same Mega Man as X. If they are the same, Mega Man underwent a major temperament change. note Some may argue that the melancholic side to him simply got brought to clearer light as Mega Man got infused with human feelings.
Among the Four Guardians: The encouraging but Trigger Happy Fefnir (sanguine/choleric), the loyal, confrontational Phantom (choleric/melancholic), the calm-but-playful Leviathan (phlegmatic/leukine), and the reliable and civil Harpuia (melancholic/sanguine).
A duplicate was made of X, but not quite the same temperament, and a lot less mature.
Evil trio of Zero 3: With what little personality he's shown to have, Omega seems to be sanguine (opposite Copy-X); Weil seems to be leukine/phlegmatic.
In Temperament, Gandrayda alone fits her respective weapon element (electric — sanguine).
The Sons Of Big Boss in Metal Gear Solid: Liquid Snake (choleric), Decoy Octopus aka DARPA chief/Vulcan Raven (melancholic), Sniper Wolf (melancholic/phlegmatic), Revolver Ocelot (phlegmatic/sanguine), Psycho Mantis (sanguine/choleric).
The Dead Cell Unit in Metal Gear Solid 2 applies: Solidus Snake (choleric), Fortune (melancholic), Vamp (phlegmatic), Fatman (sanguine).
The Cobra Unit in Metal Gear Solid 3 applies: The Pain/The Fury (choleric), The Boss/The Sorrow (melancholic), The End (phlegmatic), The Fear (sanguine).
The Beauty and the Beast unit in Metal Gear Solid 4 applies: Raging Raven (choleric), Crying Wolf (melancholic), Screaming Mantis (phlegmatic), and Laughing Octopus (sanguine).
Nazi Zombies: Tank (choleric), Takeo (melancholic), Nikolai (phlegmatic), and Richtofen (sanguine).
NieR features has something of this as well. Nier (choleric), Emil (melancholic), Grimoire Weiss (phlegmatic), and Kaine (sanguine).
Judging by their programmed natures, the Pac-Man ghosts:
Choleric and melancholic (rational): Pinky (chases and ambushes)
Melancholic and sanguine (idealist): Inky (cautious)
Sanguine and choleric (artisan): Blinky (cocky, always chases)
Leukine/phlegmatic: Clyde (indifferent and random)
Intentionally invoked in Planescape: Torment. In the end game, you meet up with three copies of yourself, who are the good incarnation (melancholic), crazy incarnation (sanguine) and pragmatic incarnation (choleric). The character of the protagonist depends on how you've been playing him, but he does start out as phlegmatic.
Amy/Big/Charmy are Sanguine being the most optimistic and fun-loving of characters.
Star Wars: Republic Commando: The four Clone Commandos of Delta Squad are this. Delta-38, "Boss", is choleric, showing good leadership instincts. Delta-07, "Sev", is melancholic, being the Cold Sniper of the group and showing great care to place each round for maximum effenciency. Delta-40, "Fixer", is phlegmatic, being, as Taun We put it, "A pure and uncomplicated soldier." Delta-62, "Scorch", is sanguine, showing much enjoyment in blowing the CIS forces apart.
In Street Fighter II, the four members of Shadaloo: Balrog (choleric), Sagat (melancholic), M. Bison (phlegmatic), and Vega (sanguine).
In the Super Mario Bros. series and its Wario Land spinoff: Mario (melancholic/phlegmatic), Yoshi and Daisy (sanguine), Luigi (sanguine/choleric), Bowser (choleric), Wario (choleric/melancholic), Toad (melancholic), and Peach and Waluigi (leukine), also Rosalina (phlegmatic).
The Koopalings and Bowser Jr.: Larry (choleric/phlegmatic), Roy (phlegmatic), Lemmy and Iggy (sanguine); Wendy and Bowser Jr. (choleric); Morton (leukine) and Ludwig (melancholic).
The five party members in Super Mario RPG: Mario (melancholic), Mallow (phlegmatic/leukine), Geno (leukine), Bowser (choleric) and Princess Toadstool (phlegmatic)
The major Paper Mario antagonists: Bowser (choleric), Sir Grodus (melancholic/choleric), Shadow Queen (leukine/phlegmatic), Count Bleck (melancholic)
Mario (melancholic), Luigi (sanguine/choleric), Starlow (sanguine), Prince Dreambert (leukine)
Big Massif and his Hooraw disciples: Sorrow Fist (melancholic), Beef Cloud (phlegmatic), Heavy Zest (sanguine), Thunder Sass (choleric) and Big Massif himself (leukine).
The ATX Team in Super Robot Wars. At first the lineup is Sanger (choleric), Bullet (melancholic), Kyosuke (phlegmatic), and Excellen (sanguine). Later, Kusuha was drafted into the team as the leukine. Later on, Sanger leaves, and Kyosuke takes on the choleric mantle; and when Lamia enters, she takes on the phlegmatic mantle.
Choleric: Fox, Ness, Bowser, King Dedede, Captain Falcon
Choleric and melancholic: Wario, Wolf, Snake
Melancholic and sanguine: Mario, Meta Knight, Marth
Sanguine: Kirby, Yoshi, Jigglypuff
Sanguine and choleric: Luigi, Donkey Kong, Pikachu, Falco, Ike
Phlegmatic/leukine: Peach, Link, Diddy Kong, Samus, Pit, Game and Watch, R.O.B., Pokemon Trainer
The Colour Tuesday: Alex is the hotheaded driver of the story (choleric). Kyle is the determined and selfless (melancholic). Bella is a slightly airy Cloudcuckoolander (sanguine). Chadd, the last permanent cast member, is a born follower (phlegmatic).
The Team Fortress 2 classes can be seen throughout the spectrum: Scout (choleric), Soldier (sanguine-choleric), Pyro (sanguine), Demoman (choleric-melancholic), Heavy (leukine, leaning choleric), Engineer (phlegmatic), Medic (phlegmatic-sanguine), Sniper (melancholic), Spy (melancholic-phlegmatic)