One-step immunoaffinity purification of bioactive human recombinant IL-1 beta with a monoclonal antibody directed to a well-exposed domain of the protein.
A monoclonal antibody (mAb) specific for human recombinant IL-1 beta (hu rIL-1 beta) was produced by immunizing BALB/c mice with hu rIL-1 beta purified with classical methods. This mAb recognizes an epitope within the highly hydrophylic fragment spanning amino acid 133-147. The affinity constant of this mAb towards IL-1 beta was determined by RIA. An affinity column was prepared by covalent binding of the mAb to Sepharose CL-4B. The column was capable of selectively binding hu rIL-1 beta produced in Escherichia coli directly from crude homogenates. The IL-1 beta protein yield was higher than 90% with a very good recovery of IL-1 beta biological activity. Moreover, the immunosorbent retained at least two thirds of its IL-1 beta-binding capacity after 20 cycles of purification.
Bigio
M
M
Sclavo Research Center, Siena, Italy.
Rossi
R
R
Borri
M G
MG
Casagli
M C
MC
Nucci
D
D
Baldari
C
C
Volpini
G
G
Boraschi
D
D
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D000939:Epitopes / Q000032:analysis; D005260:Female; D007375:Interleukin-1 / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000302:isolation & purification
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007375:Interleukin-1; D011994:Recombinant Proteins