Abstract: This chapter will detail the requirements of modern detonation nanodiamonds (DNDs) intended for biomedical applications, beginning with DND material preparations and followed by bio-related applications developed at International Technology Center. DNDs are one of the most commercially promising nanodiamonds with a primary particle size of 4–5 nm, produced by detonation of carbon-containing explosives. The structural diversity of DNDs will be described, which depend upon synthesis conditions, postsynthesis processes, and modifications. Bioapplications reviewed include ballistic delivery of bio-functionalized DND into cells, photoluminescent biolabeling, biotarget capturing and collection by electrophoretic manipulation of DNDs, and health care applications. DNDs are advantageous when compared with the other types of nanoparticles due to DND large scale synthesis, small primary particle size, facile surface functionalization, stable photoluminescence as well as biocompatibility. Currently, biotechnology applications have shown that NDs can be used for bioanalytical purposes such as protein purification or fluorescent biolabeling, while research is in the developing stages for DNDs applied as diagnostic probes, delivery vehicles, enterosorbents and advanced medical device applications.