A rapid Percoll technique for the purification of human basophils

We have refined a rapid single-step Percoll technique to obtain a basophil-enriched mononuclear cell fraction suitable for further purification. Greater than 75% of the total blood basophils were recovered from a blood-Percoll interface at a purity of between 5 and 23%. The contaminating cells were principally lymphocytes with a smaller (20-25%) percentage of monocytes. Further purification using the penicillin affinity column led to the recovery of between 28 and 64% of the total blood basophils at purities ranging from 52 to 92%, a substantial improvement over our earlier protocol. There was also a marked increase in the recovery of basophils from the affinity column over our previous technique, which lead to a moderate (5-10%) increase in overall recovery.