Genetic immunization for the recovery and purification of recombinant proteins.
A system that uses genetic immunization for recombinant protein recovery and purification is described. The genetic sequence encoding a target protein is subcloned into both a eukaryotic and a prokaryotic vector. With the eukaryotic construct, a rabbit is genetically immunized and specific polyclonal antibodies to the encoded protein raised. The prokaryotic construct is used for bacterial transformation and expression of recombinant protein. Recovery and purification of target recombinant protein are obtained by passing the lysate of expressing bacteria through an immunoaffinity column prepared with the polyclonal antibodies raised in the genetically immunized animal. This method allows purification of recombinant protein without fusion tails and can be applied to purify any protein whose encoding genetic sequence is known.
La Cava
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Department of Medicine, University of California, San Diego, La Jolla, California, 92093, USA. lacava@scripps.edu
Albani
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S
D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology*; D007106:Immune Sera / Q000096:biosynthesis; D007114:Immunization; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D014169:Transformation, Bacterial; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
C096364:BALF2 protein, Human herpesvirus 4; D004268:DNA-Binding Proteins; D007106:Immune Sera; D011994:Recombinant Proteins; D014764:Viral Proteins