Good manufacturing practice production of human stem cells for somatic cell and gene therapy.

Peripheral blood stem cells (PBSC) are used for transplantation to reconstitute the hematopoietic system after high-dose chemotherapy. PBSC are harvested from peripheral blood upon successful mobilization by cytokines and/or chemotherapy. Further in vitro manipulation steps like enrichment of CD34+ PBSC or gene transfer can be performed.

Isolation and sequencing of alpha-tubulin peptides from myxamoebae of the slime mould Physarum polycephalum.

Starting from only 5.9 mg of alpha-tubulin from myxamoebae of the slime mould Physarum polycephalum, we have isolated and sequenced peptides that account for 96% of the complete sequence. The peptides were generated by digestion of alpha-tubulin with trypsin, Staphylococcus aureus protease and cyanogen bromide.

Amino acid sequence data of alpha-tubulin from myxamoebae of Physarum polycephalum.

About 96% of the amino acid sequence of an alpha-tubulin from the slime mould Physarum polycephalum has been determined. Of 430 sequenced amino acids, 30 differ from the deduced amino acid sequence of a recently published alpha-tubulin complementary DNA from the plasmodial form of P. polycephalum.

Amino-acid sequence data of beta-tubulin from Physarum polycephalum myxamoebae.

Starting with 7.7 mg of a beta-tubulin isolated from myxamoebae of the slime mould Physarum polycephalum, 90% of the sequence has been determined by the Edman degradation of peptides generated by cyanogen bromide, trypsin and Staphylococcus aureus protease. Differences to other beta-tubulins are mainly conservative and spread evenly throughout the chain except for a high concentration at the C-terminus.