Protein aggregation particularly the formation of β-sheet rich structures called amyloid fibrils has been linked to a wide range of diseases. One such case involves the amyloid fibril formation of β2-microglobulin (β2m) and its accumulation in various tissue, leading to in vivo dialysis-related amyloidosis (DRA). D76N, a natural amyloidogenic variant of human β2m, can form amyloid fibrils in vitro within a few hours under physiological conditions. This makes β2m an appropriate model for DRA related studies. Clusterin and αB-crystallin are known to act as molecular chaperones, preventing both amorphous and amyloid aggregation. Significant amounts of clusterin and αB-crystallin are present in the renal tissue. Several reports suggest overproduction of clusterin during renal injury, this prompts us to examine the role played by clusterin and αB-crystallin
during fibrillogenesis and DRA.

In vitro investigation via Thioflavin T (ThT) binding suggests that both clusterin and αB-crystallin
prolongs the lag phase of D76N β2m amyloid formation. We also found that once the prefibrillar species are generated, both the chaperones become incorporated into the fibril formation pathway. Interestingly the amyloid like structure formed in the presence of chaperones also slows down the fibrillar conversion of D76N via amyloid seeds. Furthermore, 8-Anilinonaphthalene-1-sulfonic acid (ANS) binding assays and isothermal titration calorimetry (ITC) indicate the involvement of hydrogen bonds and hydrophobic interaction in prolonging the lag phase.