Development of an enzyme-linked immunoreceptor assay (ELIRA) for quantification of the biological activity of recombinant human bone morphogenetic protein-2

Human bone morphogenetic protein-2 is a representative of the transforming growth factor-β (TGF-β) superfamily of cytokines. It was produced in high-cell-density cultivations of recombinant Escherichia coli leading to the formation of inclusion bodies with aggregated inactive protein so that the pro...

Full description

Saved in:
Bibliographic Details
Published in:Journal of biotechnology 2005-10, Vol.119 (4), p.425-435
Main Authors: Wendler, Janine, Hoffmann, Andrea, Gross, Gerhard, Weich, Herbert A., Bilitewski, Ursula
Format: Article
Language:English
Subjects:
Biological activity
Biological and medical sciences
Biotechnology
BMPR-IA
BMPR-IB
Bone Morphogenetic Protein 2
Bone Morphogenetic Proteins - analysis
Bone Morphogenetic Proteins - genetics
Bone Morphogenetic Proteins - metabolism
Enzyme-linked immunoreceptor assay
Enzyme-Linked Immunosorbent Assay - methods
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Fundamental and applied biological sciences. Psychology
Humans
Recombinant bone morphogenetic protein-2
Recombinant Proteins
Transforming Growth Factor beta - analysis
Transforming Growth Factor beta - genetics
Transforming Growth Factor beta - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Human bone morphogenetic protein-2 is a representative of the transforming growth factor-β (TGF-β) superfamily of cytokines. It was produced in high-cell-density cultivations of recombinant Escherichia coli leading to the formation of inclusion bodies with aggregated inactive protein so that the protein had to be solubilized and renatured. Thus, the biological activity of the recombinant protein had to be determined. To avoid time-consuming cell-based assays or radioactive labelling of proteins enzyme-linked immunoreceptor assays were developed. They were based on the specific interaction between the biologically active protein and its receptors, of which the extracellular ligand binding domains were tagged with the Fc part of human IgG and expressed in insect cells. The amount of bound ligand, corresponding to the biologically active recombinant protein, was determined via enzyme-labelled antibodies. Application to various batches of protein showed that not only the amount of active protein could be quantified but also the quality of the protein preparations could be evaluated in significantly shorter analysis times than with conventional cell-based assays.
ISSN:0168-1656
1873-4863
DOI:10.1016/j.jbiotec.2005.04.021