Figure 3.

The reaction mixture of O-SP adipic hydrazide, HSA, and 0.1 M 1-ethyl-3-dimethylaminopropylcarbodiimide at room temperature for 4 hours with pH maintained at 5.25 was dialyzed against 0.2 M NaCl for 2 days at 4–7°C and was applied to a 1- × 95-cm column of CL-6B Sepharose column in 0.2 M NaCl, and 3.5-ml fractions were collected. Saccharide was determined by the anthrone reaction (14), and protein was determined by A₂₈₀. Void volume fractions were taken as conjugate: O-SP–adipic hydrazide and HSA had K_ds of 0.4 and 0.5, respectively.