Immunodiagnostic assays using reducing agents
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United States of America Patent
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May 3, 2011
Grant Date -
Nov 23, 2006
app pub date -
Aug 2, 2006
filing date -
Apr 17, 1998
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Abstract
The present invention relates to a solid phase immunoassay comprising on said solid phase an antigen in the presence of a reducing agent. The present invention also relates to a method for purifying a cysteine containing recombinantly expressed protein comprising at least 2, preferably 3 or 4 and even more preferably all of the following steps: (a) sulphonation of a lysate from recombinant host cells or lysis of recombinant host cells in the presence of guanidinium chloride followed by a subsequent sulphonation of the cell lysate, (b) treatment with a zwitterionic detergent, preferably after removal of the cell debris, (c) purification of the sulphonated version of the recombinant protein or purification of the sulphonated version of the recombinant protein with subsequent removal of the zwitterionic detergent, with said purification being preferably chromatography, more preferably a Ni-IMAC chromatography with said recombinant protein being a His-tagged recombinant protein, (d) desulphonation of the sulphonated version of the recombinant protein, preferably with a molar excess of DTT, (e) storage in the presence of a molar excess of DTT. The present invention also relates to novel HCV NS3 sequences as depicted in FIGS. 1-8.
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