Eukaryotic cell-free protein expression system especially suitable for disulfide-bridges comprising proteins

摘要

Preparing monomeric or dimeric proteins or peptides containing internal or external disulfide bonds comprise (a) providing a cell-free lysate obtained from eukaryotic cells containing functional microsomal vesicles; (b) adding a nucleic acid to the lysate; (c) maintaining the lysate with the nucleic acid for a pre-determined time at 20-35[deg] C, where the nucleic acid formed with proteins or peptides are translocated in the microsomal vesicles; and (d) dissolving the microsomal vesicles and optionally separating the obtained proteins or peptides from the lysate. Preparing monomeric or dimeric proteins or peptides containing internal or external disulfide bonds comprise (a) providing a cell-free lysate obtained from eukaryotic cells containing functional microsomal vesicles; (b) adding a nucleic acid to the lysate, where the nucleic acid encodes the protein or peptide and further comprises a signal sequence; (c) maintaining the lysate with the nucleic acid for a pre-determined time at 20-35[deg] C, where the nucleic acid formed with proteins or peptides are translocated in the microsomal vesicles; and (d) dissolving the microsomal vesicles and optionally separating the obtained proteins or peptides from the lysate. An independent claim is included for proteins, peptides or a composition containing proteins or peptides obtained by the above method.