Production of recombinant human beta2-microglobulin for scintigraphic diagnosis of amyloidosis in uremia and hemodialysis

Eur J Biochem. 2000 Feb;267(3):627-33. doi: 10.1046/j.1432-1327.2000.01071.x.


Amyloid of beta2-microglobulin (beta2m) origin can be diagnosed using 131I-radiolabelled-beta2m scintigraphy in patients with uremia and hemodialysis treatment. As the tracer beta2m is isolated from another patient, it carries the common risks, including viral infections such as Hepatitis B, C and HIV, which are associated with human plasma products. In order to exclude these risks we have produced recombinant human beta2m (rhbeta2m) in Escherichia coli. The expression vector pASK40DeltaLbeta2m(His)5 contains a C-terminal (His)5-tag for purification via immobilized metal ion affinity chromatography (IMAC). Size exclusion chromatography on a Superose 12 column represents the second step of purification. The isolated rhbeta2mH5 reacted in an immunochemically identical manner to native human beta2m, and showed a single band of approximately 11.8 kDa in Western blot analysis and revealed a single spot in two-dimensional gel electrophoresis. Mass spectrometry analysis revealed a single peak at the expected molecular mass of 12 415.8 Da. Uniformity was further proven by crystallization and N-terminal amino-acid sequence analysis. The rhbeta2mH5 protein was then produced under conditions that allow the intravenous use in humans. Intraveneously applied indium-111-labelled rhbeta2mH5 was monitored in hemodialysed patients with and without known beta2m-amyloidosis. The tracer was localized specifically to particular areas known to contain amyloid. Thus, this rhbeta2mH5 preparation is suitable for detecting amyloid-containing organs of the beta2m-class in vivo and fulfils the requirements of a tracer for common use. Finally, the use of indium-111 instead of iodine-131 has reduced the radioactive load and resulted in higher resolution.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloidosis / complications
  • Amyloidosis / diagnostic imaging*
  • Base Sequence
  • Chromatography, Affinity
  • DNA Primers / genetics
  • Escherichia coli / genetics
  • Humans
  • Indium Radioisotopes
  • Iodine Radioisotopes
  • Male
  • Middle Aged
  • Molecular Weight
  • Pentetic Acid / analogs & derivatives
  • Quality Control
  • Radionuclide Imaging
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Renal Dialysis*
  • Uremia / complications
  • Uremia / diagnostic imaging*
  • beta 2-Microglobulin / biosynthesis*
  • beta 2-Microglobulin / genetics


  • DNA Primers
  • Indium Radioisotopes
  • Iodine Radioisotopes
  • Recombinant Proteins
  • beta 2-Microglobulin
  • Pentetic Acid