FPLC size exclusion of FN-AFFUD or FN-AFBbk32 complex.

AFFUD (A) or AFBbk32 (B), 40nM, was mixed with 400nM FN for 30 min. The complex was loaded on a Superose 6 10/300 size-exclusion column and separated on FPLC. 400 nM FN (C), 40 nM AFFUD (D), or 40 nM AFBbk32 (E) alone was loaded on the same column. Absorbance at 280nm (A^280nm), and emission at 535 nm upon excitation at 485 nm (E^535nm) were measured to estimate, respectively, concentration of FN and AFpolypeptide and expressed relative to the values in starting material. Data are representative of three experiments.