Aurier has been on the sidelines since March after limping off in Cote d'Ivoire's 3-0 win over Rwanda in their final 2019 Africa Cup of Nations qualifying match.
His last outing for Spurs was during their 1-0 win against Borussia Dortmund when the London club marched through to the quarter-finals of the Champions League.
This season, Aurier has struggled for form in Mauricio Pochettino's team, making just eight Premier League appearances so far.
Ahead of the crucial fixture against Ajax at the Johan Cruyff Arena, they aim to bounce back from a 1-0 deficit from the first leg. Pochettino disclosed that the Ivory Coast international is one of the notable names that did not travel with the rest of the team to Amsterdam.
“Lamela is okay, he will train today. Serge [Aurier] is ok, but he’s not here,” Pochettino was quoted as saying by Football London.
Centre-back Davinson Sanchez and Harry Winks are also out of Wednesday’s fixture due to their respective injuries.
Aurier will hope to return to action for Tottenham Hotspur in their last Premier League game of the season, against Everton on Sunday.
Next month, the 26-year-old is expected to play a key role for Ivory Coast at the 2019 Africa Cup of Nations in Egypt.