Optimized bacterial expression of a synthetic BRIL antibody

Cooper, B.F.; Isom, G.L.

doi:10.1107/S2053230X26001548

Figure 1
BAG2 co-purifies with a 15 kDa protein which competes for BRIL binding. (a) SDS–PAGE of Protein L elution fractions following BAG2 overexpression in BL21-Gold indicating co-purification of a 15 kDa protein, later confirmed as cytochrome b₅₆₂. (b) Cation-exchange trace of pooled Protein L elution fractions following BAG2 overexpression in BL21-Gold. (c) SDS–PAGE of cation-exchange fractions following BAG2 overexpression in BL21-Gold, indicating that BAG2 maintains a stable complex with the 15 kDa protein later identified as cytochrome b₅₆₂ throughout purification. (d) Region of interest from a size-exclusion chromatography trace following incubation of a BRIL fusion protein (∼37 kDa) with BAG2 purified from BL21-Gold indicating two distinct complexes with different retention volumes. The full trace, highlighting the region of interest, is provided as an inset at the top right. (e) SDS–PAGE of size-exclusion chromatography fractions following incubation of a BRIL fusion protein (∼37 kDa) with BAG2 purified from BL21-Gold. Fractions corresponding to complex 1 comprise the expected BRIL fusion protein–BAG2 complex, whilst complex 2 fractions comprise BAG2 in complex with the 15 kDa protein later identified as cytochrome b₅₆₂.

STRUCTURAL BIOLOGY
COMMUNICATIONS

ISSN: 2053-230X

Volume 82| Part 4| April 2026| Pages 143-149

https://doi.org/10.1107/S2053230X26001548

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