Biotinylated liposomes can be conjugated noncovalently with (strept)avidin through either direct interaction with the protein/antibody conjugated to (strept)avidin or by coupling with other biotinylated proteins using (strept)avidin as a bridging molecule. Both avidin and (strept)avidin form strong non-covalent bond with biotin. The high resistance to breakdown makes them very useful in bioconjugate chemistry. However, (strept)avidin has replaced avidin in most bio-conjugation applications due to its enhanced properties. NeutrAvidin (ThermoFisher) is a modified avidin without negative properties. It performs much better than original avidin and sometimes(strept)avidin. In order to exploit the high-affinity interaction of biotin with (strept)avidin, a two-step “sandwich” protocol (Method A) has been developed for the preparation of targeted immunoliposomes. In this methodology, (strept)avidin is first attached to biotinylated liposomes, then a biotin-modified protein/antibody is introduced into the biotinylated (strept)avidin-labeled liposomes. This non-covalent approach is rapid, extremely versatile and applicable to numerous targeting ligands of interest with respect to in vitro and in vivo applications. Alternatively, instead of forming a (strept)avidin bridge, (strept)avidin molecule can also be covalently conjugated to antibody or ligand (Method B) and non-covalently bound to liposomes containing biotin on surface in order to form immunoliposomes.