PRODUCTION OF RECOMBINANT HUMAN LYSOSOMAL ALPHA-MANNOSIDASE

摘要

The present invention relates to a cell capable of producing recombinant human LAMAN (rhLAMAN), said cell comprising the 3066 basepair EcoRI - XbaI fragment of a human cDNA which codes for a human LAMAN protein in which the position corresponding to position 186 of the full length hLAMAN protein is Aspartic acid (Asp, D). In particular, the invention relates to a cell comprising a DNA sequence which codes for the amino acid sequence shown in SEQ ID NO. 12. One embodiment is an expression plasmid pLamanExp1 having the sequence shown in SEQ ID NO. 2. One aspect of the invention relates to a method for the preparation of recombinant human LAMAN, the method comprising a) introducing, into a suitable vector, a nucleic acid fragment comprising a DNA fragment which codes for the amino acid sequence shown in SEQ ID NO. 12, b) transforming a cell with the vector obtained in step a), c) culturing the transformed host cell under conditions facilitating expression of the nucleic acid sequence, d) recovering the expression product from the culture. The method may further comprise a fermentation step and/or a purification step. The invention further relates to a rhLAMAN produced by the method of the invention and to the use of the rhLAMAN produced according to the invention for the preparation of a medicament for the treatment of alfa mannosidosis.