One-step immunoadsorbent column purification of interleukin-4 from murine D10 supernatants.

A one-step purification of interleukin-4 is described using an 11B11 monoclonal antibody-Sepharose 4B chromatography column. Beginning with 1,300 ml of supernatant from the murine T-cell clone D10, a homogeneous preparation of IL-4 is obtained (22 micrograms) having a specific activity of 4.75 x 10(6) units/mg (10,650-fold purification) with an overall yield of 45%. The purified protein runs as a single band on silver-stained SDS polyacrylamide gels with a molecular mass of 18,600 +/- 1,000 daltons. Digestion with endoglycosidase F reduces the molecular mass to 15,500 daltons, indicating the presence of N-linked glycosylation. The novelty in this procedure involves the use of native conditions throughout and the absence of a requirement for HPLC resolution. Furthermore, the use of these cells (D10), rather than EL4 cells which have previously been used as a source of IL-4, may facilitate purification since D10 can be stimulated under serum-free conditions.