Transfer to a nitrocellulose membrane allows of the bone forming activity of bone morphogenetic protein

When we purify bone morphogenetic protein (BMP), its activity diminishes, and the quantity we are able to extract decreases. It is difficult to evaluate the effects of each of the processes involved in BMP purification because it is unstable. In order to resolve the problem, a modified bioassay method using only slight quantities of BMP which do not decrease its bone-forming activity, is needed. We transferred BMP separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) onto nitrocellulose membranes (NC) and cut out the bands. Then we used each band as implanted material. The NC membrane firmly bound the BMP activity fraction and held it in the implantation area of the mouse thigh. No formation of bone-like tissue was detected histologically at 14 days after the implantation, but, by 21 days after implantation, cartilage like tissue had clearly formed and newly formed bone was seen by 28 days. By implanting BMP transferred to individual NC membranes, we could perform a bioassay easily with small amounts of BMP without any reduction in activity.