Sharpless originally planned to attend medical school after his undergraduate degree, but his research professor convinced him to continue his education in chemistry.

[9] Sharpless developed stereoselective oxidation reactions, and showed that the formation of an inhibitor with femtomolar potency can be catalyzed by the enzyme acetylcholinesterase, beginning with an azide and an alkyne.

[11][2] This involves a set of highly selective, exothermic reactions which occur under mild conditions; the most successful example is the azide alkyne Huisgen cycloaddition to form 1,2,3-triazoles.

[1][2] He was awarded the 2001 Wolf Prize in Chemistry together with Henri B. Kagan and Ryoji Noyori “for their pioneering, creative and crucial work in developing asymmetric catalysis for the synthesis of chiral molecules, greatly increasing humankind´s ability to create new products of fundamental and practical importance”.

In 2019, Sharpless was awarded the Priestley medal, the American Chemical Society's highest honor, for "the invention of catalytic, asymmetric oxidation methods, the concept of click chemistry and development of the copper-catalyzed version of the azide-acetylene cycloaddition reaction.".

[10] He was blinded in one eye during a lab accident in 1970 where an NMR tube exploded, shortly after he arrived at MIT as an assistant professor.

After this accident, Sharpless stresses "there's simply never an adequate excuse for not wearing safety glasses in the laboratory at all times.