Single-step monoclonal antibody affinity purification of human urogastrone from urine.

The use of a monoclonal antibody (MAb) affinity column to purify epidermal growth factor/urogastrone from human urine in a single-step process is described. The MAb was raised against purified recombinant human urogastrone derived from the expression of a cloned synthetic urogastrone gene. The MAb was characterised and shown to cross-react fully with recombinant and native human urogastrones. The material eluted from the column was shown to have retained biological activity. When the eluted material was run on SDS/PAGE under reducing conditions an apparently homogeneous species of 5400 Da apparent molecular weight was seen. When examined on acid PAGE, 2 molecular species corresponding to beta and gamma urogastrone were observed.