Having started brightly with a goal chalked off for offside inside the opening minute, the Glovers found themselves behind after 20 minutes.
Uche Ikpeazu broke the offside trap to leave the Yeovil defence appealing, but the linesman’s flag didn’t come as the U’s top scorer round Artur Krysiak to score.
Buoyed by the opener it was 2-0 soon after as George Maris punished a lapse in concentration to put the home side well and truly in the driving seat.
Defender Bevis Mugabi reduced the arrears on 59 minutes to set up a final half-hour with all to play for.
The visitors came close to an equaliser in the closing stages, but United held on for all three points to leave the Green and Whites deflated.
Darren Way was forced into making one change from the side that saw off Crewe Alexandra at Huish Park on Saturday.
Picking up his fifth booking of the season against the Railwaymen, Francois Zoko served a one-game suspension allowing young loanee Sam Surridge to start upfront.
Sid Nelson, who missed his first match on the weekend since joining on loan, took a seat amongst the substitutes in Cambridgeshire after recovering from surgery on a broken nose in the only other change to the matchday squad.
Oozing with confidence from collecting three points just three days ago, Town had the ball in the back of the net after just 30 seconds only for the linesman’s flag to rule out a dream start.
A clever kick-off routine saw a free-flowing football manoeuvre end with Olufela Olomola firing past David Forde, only for Sam Surridge to be pulled up for offside having followed in Rhys Browne’s low drive which started the flurry of shots.
The early signs were encouraging for the Green and Whites, but the warning signs emerged with 10 minutes played.
A perfectly timed Bevis Mugabi challenge stopped Jabo Ibehre calling Krysiak into action from the angle, before the Polish stopper was left blameless for the opener.
Having dealt with copious balls into the box the visitors’ offside trap was sprung as Jevani Brown played in Ikpeazu, who coolly rounded Krysiak to make it 1-0.
In a costly spell for Town, Shaun Derry’s side doubled their advantage just six minutes later, as Bradley Halliday’s teasing cross picked out Maris who stole in unmarked from midfield to head home.
Punished for a 10-minute wobble, the Glovers recovered to enjoy the lion share of possession up to the break.
However, unable to draw Forde into meaningful action before the referee’s whistle ended first-half proceedings, Way’s side trudged into the dressing room facing a two-goal deficit.
With Cambridge starting on the front foot after the restart, Way wasted no time in making a change as the attacking attributes of Jake Gray were introduced in place of Connor Smith.
Met with a change in shape which saw real fluidity introduced amongst the attacking ranks, Town found a way back into the match just before the hour-mark.
Committing numbers forward, Ryan Dickson’s pinpoint delivery picked out a Green and White shirt in the middle for the second match in a row, as Mugabi – up from right-back – rose highest to head powerfully into the bottom corner.
Deservedly with a foothold back in the match, Yeovil were almost punished for a sense of deja vu.
Halliday’s ball into the middle once again picked out an unmarked Maris, but the U’s midfielder could only direct a header off the post for a goal-kick as the final 20 minutes loomed.
Becoming an end-to-end affair, Way opted to go for broke by introducing Alefe Santos for captain James Bailey, desperate to come away with something to show for an encouraging performance.
Inside the final five minutes an important Leon Legge header stopped Surridge’s cross finding Olomola at the back post, before Otis Khan saw a low drive cleared off the line by Ibehre as Town threw everything at Cambridge.
Despite a valiant effort it wasn’t enough to come away from the Abbey Stadium with a point, as a quickfire double in the first-half for United condemned Yeovil to defeat.
Cambridge United: Forde, Halliday, Legge, Taylor, Carroll, Deegan, Mingoia, Maris (Elito 87), Brown, Ibehre, Ikpeazu (O’Neil 90).
Subs not used: Mitov, Dunk, Taft, Howkins, Foy.
Yeovil Town: Krysiak, Mugabi, Sowunmi, N. Smith, Browne, Bailey (Santos 81), C. Smith (Gray 51), Khan, Olomola, Surridge.
Subs not used: Maddison, Nelson, Davies, James, Worthington.