The production of a recombinant tandem single chain fragment variable capable of binding prolamins triggering celiac disease

BMC Biotechnology
Britta EggenreichOliver Spadiut

Abstract

Celiac disease (CD) is one of the most common food-related chronic disorders. It is mediated by the dietary consumption of prolamins, which are storage proteins of different grains. So far, no therapy exists and patients are bound to maintain a lifelong diet to avoid symptoms and long-term complications. To support those patients we developed a tandem single chain Fragment variable (tscFv) acting as a neutralizing agent against prolamins. We recombinantly produced this molecule in E. coli, but mainly obtained misfolded product aggregates, so-called inclusion bodies, independent of the cultivation strategy we applied. In this study, we introduce this novel tscFv against CD and present our strategy of obtaining active product from inclusion bodies. The refolded tscFv shows binding capabilities towards all tested CD-triggering grains. Compared to a standard polyclonal anti-PT-gliadin-IgY, the tscFv displays a slightly reduced affinity towards digested gliadin, but an additional affinity towards prolamins of barley. The high binding specificity of tscFv towards prolamin-containing grains makes this novel molecule a valuable candidate to support patients suffering from CD in the future.

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Citations

Jul 4, 2019·Bioprocess and Biosystems Engineering·Alexander PekarskyOliver Spadiut
Jun 4, 2019·Frontiers in Pediatrics·Shakira Yoosuf, Govind K Makharia
Jan 1, 2020·Journal of Biotechnology·Britta EggenreichOliver Spadiut

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Methods Mentioned

BETA
Enzyme-linked immunosorbent assay
ELISA
ELISAs

Clinical Trials Mentioned

NCT02472119
NCT01990885
NCT01255696
NCT00962182
NCT00810654
NCT01257620
NCT01661933
NCT02754609
NCT01396213
NCT01893775

Software Mentioned

OriginPro
GPM
SigmaPlot

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