Erythropoietin (EP) is a primary regulator of erythropoiesis. Accordingly, its possible diagnostic and treatment applications in certain refractory anemias has been postulated. The lack of both raw material for erythropoietin purification and relatively pure EP (150 units per milligram of protein) has severely compromised its use in research, diagnosis, and treatment. This situation has been complicated by the lack of an accurate, simple assay for biologically active EP. The long-term objectives of this investigation are the development of economical procedures for the generation, assay, and purification of erythropoietin.Using sheep and a novel blood plasmapheresis procedure, it is believed that large volumes of EP-rich plasma (2-10 units per milliliter) can be economically produced. This approach will allow for a low-cost supply of plasma for EP assay and purification. The in vitro plasma clot technique for culturing erythroid progenitors will be refined and developed as an erythropoietin bioassay. Purification will initially rely on a simplified partial purification procedure based on nitrocellulose-EP binding affinity and standard purification procedures to yield a pure enough EP antigen to generate highly specific F-P-antisera (polygonal and/or monoclonal). This could in turn be used to efficiently purify erythropoietin.Institute Of Arthritis, Diabetes, And Digestive And Kidney Diseases
