Iodoacetic Acid → HCID 131726016, compound

Participants

Reaction Steps

To one volume of a solution of an Fab fragment (the same solution conditions as that used in (a) above) were added two volumes of a solution of 1M p-aminophenylalanine amide (pH adjusted to pH 6.7 with acetic acid). Thre volumes of butane-1,4-diol were added, followed by 0.06 volumes of cysteine (1M). A 50:50 mixture of papain suspension and butane-1,4-diol was prepared (final papain content 18 mg/ml) and 0.28 volumes of the mixture were added at once to the Fab solution. The apparent pH was checked with pH paper: if the apparent value was lower than pH 6.2 it was raised to 6.2–6.7 with 0.01M NaOH (with careful, rapid mixing). After 18 hours at room temperature the papain was inactivated by addition of the appropriate volume of iodoacetic acid, 0.5M, brought to pH 7.0 with 1% NaHCO3 solution. After incubation for 10 minutes at room temperature the protein conjugate was isolated on Sephadex G50 as described above.

Provenance & Evidence