Methods for preparing human thrombopoietin polypeptides by mammalian cell cultures

摘要

An expression vector replicable in mammalian cells contains: i) a hTPO transcription portion consisting of: a) a transcription promoter derived from a variable kappa gene of human immunoglobulins; b) a DNA segment encoding the complete hTPO polypeptide including its signal peptide; c) an immunoglobulin transcription enhancer DNA segment of the human kappa gene; d) a transcription terminator; and e) a polyadenylation signal (ii) a gene encoding resistance to the antibiotic neomicyn type drugs (iii) a gene encoding resistance to the antibiotic ampicillin; and (iv) a replication origin for E Coli ColEl ori. The vector allows biologically active human thrombopoietin to be obtained from in vitro cultures of mammalian cells. Preferably the TPO is obtained from a cultured cell line derived from the murine myeloma X-63 (ATCC: P3X63Ag8.6533, CRL 1580).