The 39-year-old continued to build Pakistan's lead after opener Khurram Manzoor fell for 146, taking his team to 352-4 at lunch in reply to South Africa's first innings total of 249.
At the break Misbah was unbeaten on a brilliant 78 and with him Asad Shafif was not out on 40, having added 62 for an unbroken fifth wicket stand.
Pakistan were leading by 103 runs with six wickets in hand.
Resuming at 263-3, Khurram Manzoor and Misbah saw off the second new ball confidently which left the South African pacemen frustrated as they looked for an early wicket.
But paceman Vernon Philander gave South Africa the much-needed breathrough by dismissing Manzoor in the sixth over of the day, caught off a slash in the slip by Jacques Kallis.
Manzoor hit 15 boundaries and batted solidly through his 388-minute knock. He added 112 for the fourth wicket with his skipper to strengthen Pakistan's position.
Shafiq, playing for his place in the team after a disastrous Zimbabwe tour where he managed just 43 in four innings, batted with authority. He has so far hit four boundaries in his 87-ball knock.
South Africa's lone regular spinner Robin Peterson has also failed to have any impact, conceding 111 runs without taking a wicket in 27 overs.
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