Recently, cryopreservation of AuNPs without aggregation has been attempted to improve their long-term stability. This study investigated criteria to select cryoprotectants for AuNPs using a variety of materials, including sugar (sucrose), surfactant (Tween 20), polymers (polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP)), and biopolymer (pectin). For cryoprotective performance, UV-vis spectroscopy reveals the potential of all cryoprotectants for preventing citrate-capped AuNPs (cit-AuNPs) from irreversible aggregation under freezing. While sucrose, PVP, and pectin were more suitable than Tween 20 and PVA as cryoprotectants for lyophilization of AuNPs with the maintained redispersability. For storage and further use, Luria-Bertani agar plate, dynamic light scattering (DLS), and transmission electron microscopy (TEM) results indicate impacts of the cryoprotectant coexisted with AuNPs after resuspension and imply that washing of the restored AuNPs is encouraged. Otherwise, running the restored AuNPs through applications, such as functionalization, protein conjugation, and surface-enhanced Raman scattering (SERS), without washing the cryoprotectant could lead to inaccurate results. This study also serves as a guideline for a comprehensive practice flow of AuNP handling, encompassing the synthesis step, cryopreservation, and use after resuspension.