Sharpless was born in Philadelphia. He graduated from Friends' Central School in 1959. He continued his studies at Dartmouth College (1963) and earned his Ph.D from Stanford University in 1968. He continued post-doctoral work at Stanford University and Harvard University. He holds honorary degree of Technical University of Munich.
In 2001 he won the Nobel Prize in Chemistry for his work on stereoselective oxidation reactions (Sharpless epoxidation, Sharpless asymmetric dihydroxylation, Sharpless oxyamination). This prize was shared with William S. Knowles and Ryoji Noyori (for their work on stereoselective hydrogenation). He also successfully epoxidized (using racemic tartaric acid) a C-86 Buckminster Fullerene ball, employing p-cresol as solvent. Currently he spends much of his time promoting click chemistry, a set of highly selective, exothermic reactions which occur under mild conditions; the most successful variant of which is the azide alkyne Huisgen cycloaddition to form 1,2,3-triazoles.