High level expression of birch pollen profilin (Bet v 2) in Escherichia coli: purification and characterization of the recombinant allergen.
Up to 20% of the population in industrialized countries suffer from type I allergic symptoms (rhinitis, conjunctivitis, and bronchial asthma). The cDNA coding for birch pollen profilin, a highly conserved cross-reactive allergen and actin-binding protein was expressed in Escherichia coli. Upon induction with IPTG up to 30 mg recombinant profilin per liter culture could be obtained. A single step purification protocol based on the high affinity of profilin to poly-(L-proline) Sepharose was used to obtain large amounts of soluble and pure recombinant birch profilin. Recombinant birch pollen profilin specifically bound IgE, elicited dose dependent histamine release from patients basophils and could be used for skin prick testing without toxic effects. The results indicate that by using purified recombinant profilin, specific diagnosis of type I allergy might be improved.
Susani
M
M
Institute of Molecular Biology, Academy of Science, Salzburg, Austria.
Jertschin
P
P
Dolecek
C
C
Sperr
W R
WR
Valent
P
P
Ebner
C
C
Kraft
D
D
Valenta
R
R
Scheiner
O
O
D000327:Adsorption; D000485:Allergens / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D001483:Base Sequence; D001491:Basophils / Q000276:immunology; D003285:Contractile Proteins; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006636:Histamine Release; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000097:blood; D008840:Microfilament Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010957:Plasmids; D011058:Pollen / Q000737:chemistry*; D051304:Profilins; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D012882:Skin Tests; D014197:Trees
D000485:Allergens; D003285:Contractile Proteins; D008840:Microfilament Proteins; D051304:Profilins; D011994:Recombinant Proteins; D007073:Immunoglobulin E