Hydrophobic Interaction of Human Interferon with Concanavalin A-Agarose

Mary W Davey,Jiunn W Huang,Eugene Sulkowski,William A Carter

doi:10.1016/s0021-9258(19)42261-8

Abstract

Abstract Human interferon binds to concanavalin A-agarose at high ionic strength (1.0 m NaCl) and cannot be readily displaced from the column with 0.1 m α-methyl-d-mannopyranoside. A partial recovery of interferon activity, with a protracted elution profile, was obtained with ethylene glycol, a polarity reducing agent. Complete recovery of activity, with a sharp elution profile, was obtained by the combined action of sugar and ethylene glycol. Apparently, the binding of human interferon to concanavalin A-agarose is due to both carbohydrate recognition and hydrophobic interaction.

Highlights

Human interferon binds to concanavalin A-agarose at high ionic strength (1.0 M NaCl) and cannot be readily displaced from the column with 0.1 M a-methyl-D-mannopyranoside
(1 M NaCl), there is protracted elution of interferon activity, amounting to 7 76 of that applied on the column
Rabbit interferon when applied OJI concanavalin A-agarose column was always recovered with sugar alone; this is consistent with the apparent lack of hydrophobic binding of rabbit interferon to albumin-agarosc [3]

Summary

SUMMARY

Human interferon binds to concanavalin A-agarose at high ionic strength (1.0 M NaCl) and cannot be readily displaced from the column with 0.1 M a-methyl-D-mannopyranoside. When the column is developed with eluant (El) containing 0.1 M a-methyl-n-mannopyranoside (1 M NaCl), there is protracted elution (tubes .20 to 80) of interferon activity, amounting to 7 76 of that applied on the column This seems to indicate that binding, in part, is due to carbohydrate recognition. That sugar alone is very inefficient in the displacement of bound interferon Development of another column (Fig. 1B) with ethylene glycol (50yG v/v) in the eluant (Ez), resulted in part’ial but quantitatively significant (650/,) recovery of interferon activity. This recovery of interferon activity with ethylene glycol indicates the involvement of hydrophobic interaction [8] in its binding to concanavalin A-agarose. Rabbit interferon when applied OJI concanavalin A-agarose column was always recovered with sugar alone; this is consistent with the apparent lack of hydrophobic binding of rabbit interferon to albumin-agarosc [3]

The selectivity of binding of human interferon to concanavalin

Findings

Fraction Number