A meeting with the Spanish champions provides a daunting test at the best of times, but Gerardo Martino's side are likely to be particularly focused this weekend after suffering a shock defeat in their previous league fixture.
Goals from Dani Parejo, Pablo Piatti and Paco Alcacer handed Valencia a 3-2 win at Camp Nou and condemned Barcelona to a first Liga defeat at home since April 2012.
The Catalan club bounced back on Wednesday with a 2-0 victory over Real Sociedad in the first leg of their Copa del Rey semi-final.
However, for the first time under Martino, Barca now face the challenge of reining in new league leaders.
Atletico Madrid hold a three-point lead at the summit after city rivals Real also dropped points in a 1-1 draw at Athletic Bilbao.
Forward Pedro, who is in line to make his 250th first-team appearance, underlined the importance of the match at Estadio Ramon Sanchez Pizjuan.
He told Barca's official website: "This game is of utmost importance. We have a lot to play for.
"There is very little margin for error between now and the end of the season; we practically can't afford to make any mistakes."
Barca have claimed three points from their previous two games at Sevilla and are unbeaten in eight matches against the Andalucian outfit.
However, Sevilla wasted a golden opportunity to claim victory in last season's corresponding fixture, as late goals from Cesc Fabregas and David Villa completed a dramatic comeback for the visitors, who trailed 2-0 before eventually triumphing 3-2.
An injury-time strike from Alexis Sanchez ensured Barca also triumphed 3-2 when the sides met in September, while Sevilla went down by the same margin at Malaga last weekend.
Unai Emery's men have gone four games without a win since beating Getafe on January 5, a run that has damaged their hopes of securing European qualification.
Emery will be without suspended full-back Coke, who picked up his fifth yellow card of the season at Malaga, while Stephane Mbia (knee) is a few weeks away from a return.
Javier Mascherano and Jordi Alba are banned for Barcelona, while Neymar (ankle) appears unlikely to feature.
