Figure 5. High-performance liquid chromatography–size exclusion profile of gamma ray irradiated αA-crystallin wild-type and N-terminal mutants. The high-performance liquid chromatography (HPLC)–size exclusion chromatographic (SEC) analysis of αA-crystallin wild-type (wt) and N-terminal mutants NGI (A) and gamma irradiation (GI) at a dosage of 0.5 kGy (B), 1.0 kGy (C), and 2.0 kGy (D) using the TSKgel-G4000SWXL column. D1 inset represents the distinct peaks of αA-crystallin wt and R21L GI at 2.0 kGy. An aliquot of 100 µl of 1 mg/ml protein was injected onto the column, and the elution profile was recorded by absorbance detection at 280 nm. All the non-gamma irradiation (NGI) mutants exhibited slight increased oligomeric size compared to the αA-crystallin wt. The SEC profile at 0.5 kGy shows the slight increased oligomeric size of all mutants compared to wild-type. At 1.0 kGy and 2.0 kGy, all the mutants except R21L demonstrate a notable increase in the aggregate size. Formation of high molecular weight (HMW) aggregates with the profile peak near to the void volume represents the increased aggregate size with an apparent molecular mass of >7 × 10⁶ Da. The SEC profile of αA-crystallin wt appears stable in all conditions.