“…The biotin-(strept)avidin interaction has a long history in the AFM-SMFS community (Florin et al, 1994(Florin et al, , 1995Lee et al, 1994;Moy et al, 1994;Grubmüller et al, 1996;Wong et al, 1999;Yuan et al, 2000;Pincet and Husson, 2005;de Odrowaz Piramowicz et al, 2006;Rico and Moy, 2007;Erdmann et al, 2008;Guo et al, 2008;Zhang et al, 2009;Chivers et al, 2010;Taninaka et al, 2010a,b;Teulon et al, 2011;Han et al, 2012;Rico et al, 2015Rico et al, , 2019Baumann et al, 2016;Erlich et al, 2017;Sedlak et al, 2017Sedlak et al, , 2019Sedlak et al, , 2020Bauer et al, 2018), and a thorough review of this controversy is beyond the scope of the review here. Much of the irreproducibility of biotinstreptavidin rupture force measurements by AFM-SMFS can be attributed to random lysine-based immobilization of the protein, as well as the fact that the streptavidin tetramer can disassemble during stretching.…”