PMID: 8960907Dec 1, 1996Paper

Expression of human recombinant beta 2-microglobulin by Aspergillus nidulans and its activity

Human Immunology
S M O'HerrinW J Burlingham

Abstract

The light chain of HLA class I protein (beta 2m) has been expressed in Aspergillus nidulans. The cDNA of beta 2m was modified using the polymerase chain reaction to include overlapping extensions for its subsequent fusion into an Aspergillus vector. This fusion resulted in beta 2m cDNA being flanked by the Aspergillus awamori glucoamylase promoter and the Aspergillus niger glucoamylase terminator. Expression of beta 2m was induced by the addition of starch to the culture medium. In preliminary mass culture trials, 177 micrograms/liter of f beta 2m were obtained in 60-liter fermentations. N-terminal sequencing of purified human beta 2m produced in fungi (f beta 2m) revealed that 28% of the purified protein was of proper sequence and 61% of the protein had an additional serine and lysine residue derived from the C-terminus of the fungal leader. Purified f beta 2m from culture supernatants appeared biochemically similar to beta 2m obtained from human urine (u beta 2m) as seen in immunoblot analysis. Functionally, f beta 2m effectively interacted as a subunit of class I MHC molecules. This was seen both in a sandwich ELISA for detecting properly folded HLA class I heavy chain and in assays showing cell-surface beta 2m exchange into...Continue Reading

References

Nov 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·M G MageD H Margulies
Feb 1, 1991·European Journal of Immunology·E MottezP Kourilsky
Jul 27, 1990·Cell·A TownsendA Tse
Jan 1, 1989·Annual Review of Immunology·A Townsend, H Bodmer
Apr 1, 1994·Enzyme and Microbial Technology·D A MacKenzieD B Archer
Oct 1, 1994·Protein Expression and Purification·T BrandhorstW R Kenealy

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Citations

Mar 18, 2006·Advances in Applied Microbiology·O P WardA Singh

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