TY - JOUR
T1 - Recombinant allergen Lol p II
T2 - Expression, purification and characterization
AU - Tamborini, Elena
AU - Brandazza, Anna
AU - De Lalla, Claudia
AU - Musco, Giovanna
AU - Siccardi, Antonio G.
AU - Arosio, Paolo
AU - Sidoli, Alessandro
PY - 1995
Y1 - 1995
N2 - Pollen from perennial rye grass (Lolium perenne) is a major cause of type I allergies worldwide. It contains complex mixtures of proteins, among which Lol p II is a major allergen. Previously, we have reported the cloning and sequencing of Lol p II and its expression in fusion with the heavy chain of human ferritin as carrier polypeptide (Sidoli et al. 1993, J. biol. Chem. 268, 21819-21825.). Here, we describe the expression, purification and characterization of a recombinant Lol p II overproduced as a non-fusion protein in the periplasm of E. coli. The recombinant allergen was expressed in high yields and was easily purified in milligram amounts. It competed with the natural Lol p II for binding to specific IgE, and it induced allergic responses in skin prick tests, indicating to be immunologically analogous to the natural protein. Biochemical analyses indicate that recombinant Lol p II is a highly stable and soluble monomeric molecule which behaves like a small globular protein.
AB - Pollen from perennial rye grass (Lolium perenne) is a major cause of type I allergies worldwide. It contains complex mixtures of proteins, among which Lol p II is a major allergen. Previously, we have reported the cloning and sequencing of Lol p II and its expression in fusion with the heavy chain of human ferritin as carrier polypeptide (Sidoli et al. 1993, J. biol. Chem. 268, 21819-21825.). Here, we describe the expression, purification and characterization of a recombinant Lol p II overproduced as a non-fusion protein in the periplasm of E. coli. The recombinant allergen was expressed in high yields and was easily purified in milligram amounts. It competed with the natural Lol p II for binding to specific IgE, and it induced allergic responses in skin prick tests, indicating to be immunologically analogous to the natural protein. Biochemical analyses indicate that recombinant Lol p II is a highly stable and soluble monomeric molecule which behaves like a small globular protein.
KW - allergens
KW - IgE
KW - perennial rye grass
KW - recombinant proteins
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U2 - 10.1016/0161-5890(95)00011-3
DO - 10.1016/0161-5890(95)00011-3
M3 - Article
C2 - 7783753
AN - SCOPUS:0029031586
VL - 32
SP - 505
EP - 513
JO - Molecular Immunology
JF - Molecular Immunology
SN - 0161-5890
IS - 7
ER -