Sep 29, 1989

One-step immunoaffinity purification of bioactive human recombinant IL-1 beta with a monoclonal antibody directed to a well-exposed domain of the protein

Journal of Immunological Methods
M BigioD Boraschi

Abstract

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.

Mentioned in this Paper

Monoclonal Antibodies
Covalent Interaction
Mice, Inbred BALB C
Interleukin-1
Proteins, Recombinant DNA
Antigenic Specificity
One-Step Dentin Bonding System
Sepharose CL 4B
Purification Aspects
Recombinants

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