Meeting the specified requirement will unlock the corresponding biocard.

Biocard	How to unlock
01. Robin	Beat Mission 1 on Easy difficulty or Normal difficulty.
02. Cyborg	Beat Mission 2 on Easy difficulty or Normal difficulty.
03. Raven	Beat Mission 3 on Easy difficulty or Normal difficulty.
04. Starfire	Beat Mission 4 on Easy difficulty or Normal difficulty.
05. Beast Boy	Beat Mission 5 on Easy difficulty or Normal difficulty.
06. Hive Thug	Collect all 25 Titan cards in Mission 1 on Easy difficulty or Normal difficulty.
07. Gizmo	Collect all 25 Titan cards in Mission 2 on Easy difficulty or Normal difficulty.
08. Jinx	Collect all 25 Titan cards in Mission 3 on Easy difficulty or Normal difficulty.
09. Mammoth	Collect all 25 Titan cards in Mission 4 on Easy difficulty or Normal difficulty.
10. Brother Blood (regular white attire)	Beat Mission 6 in under 25 minutes on Normal difficulty.
11. Brother Blood (cybernetic red attire)	Collect all 25 Titan cards in Mission 6 on Easy difficulty or Normal difficulty.
12. Terra	Collect all 150 Titan cards in the game on Easy difficulty or Normal difficulty.
13. Bumblebee	Beat Mission 4 with a success rate of 80% or higher on Hero difficulty.
14. Speedy	Beat Mission 1 in under 20 minutes on Hero difficulty.
15. Más y Menos	Beat Mission 5 in under 20 minutes on Titans difficulty.
16. Mad Mod	Beat Mission 2 in under 20 minutes on Normal difficulty.
17. Control Freak	Collect all 25 Titan cards in Mission 5 on Easy difficulty or Normal difficulty.
18. Puppet King	Beat Mission 4 in under 20 minutes on Normal difficulty.
19. Mumbo Jumbo	Collect all 150 Titan cards in the game on Hero difficulty.
20. Johnny Rancid	Beat Mission 5 with a success rate of 80% or higher on Hero difficulty.
21. Cinderblock	Beat Mission 6 with a success rate of 80% or higher on Titans difficulty.
22. Slade	Beat Mission 1 with a success rate of 95% or higher on Normal difficulty.
23. Larry the Titan	Collect all 150 Titan cards in the game on Titans difficulty.
24. Red X	Beat Mission 1 using only Robin on Normal difficulty.
25. Trigon	Beat the game on Titans difficulty.