Refolding of Inclusion Body Proteins from
            <i>E. coli</i>

Su, Zhiguo; Lu, Diannan; Liu, Zheng

doi:10.1002/9780470939932.ch13

Cited by 13 publications

(10 citation statements)

References 180 publications

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“…The rhSCF was expressed as IBs in E. coli . IB was formed from partially folded protein intermediates and was composed of aggregates of mostly single types of polypeptide . The IBs were isolated and washed to remove large numbers of host proteins and/or debris, and further improved the degree of rhSCF.…”

Section: Discussionmentioning

confidence: 99%

“…To obtain biologically active proteins from IB, a refolding step is required. Refolding is commonly performed after solubilization in denaturing solutions, followed by dilution . The dilution refolding method is also widely used for its simplicity to perform and scale‐up .…”

Section: Discussionmentioning

confidence: 99%

“…Pharmacological importance of recombinant human stem cell factor (rhSCF) has increased the demand to establish effective and large‐scale production and purification processes. Several protocols have been developed for the expression and purification of hSCF in E. coli , CV‐1 in Origin with SV40 genes‐7 (COS‐7) cells, and Chinese hamster ovary (CHO) cells as soluble and inactive aggregates on a laboratory scale . These protocols resulted in the production of milligram amounts of rhSCF‐used bacterial expression systems.…”

Section: Introductionmentioning

confidence: 99%

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Large‐scale purification and characterization of recombinant human stem cell factor in Escherichia coli

Chen

Cai

Zhang

et al. 2017

Biotech and App Biochem

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The pharmacological importance of recombinant human stem cell factor (rhSCF) has increased the demand to establish effective and large-scale production and purification processes. A good source of bioactive recombinant protein with capability of being scaled-up without losing activity has always been a challenge. The objectives of the study were the rapid and efficient pilot-scale expression and purification of rhSCF. The gene encoding stem cell factor (SCF) was cloned into pBV220 and transformed into Escherichia coli. The recombinant SCF was expressed and isolated using a procedure consisting of isolation of inclusion bodies (IBs), denaturation, and refolding followed by chromatographic steps toward purification. The yield of rhSCF reached 835.6 g/20 L, and the expression levels of rhSCF were about 33.9% of the total E. coli protein content. rhSCF was purified by isolation of IBs, denaturation, and refolding, followed by SP-Sepharose chromatography, Source 30 reversed-phase chromatography, and Q-Sepharose chromatography. This procedure was developed to isolate 5.5 g of rhSCF (99.5% purity) with specific activity at 0.96 × 10 IU/mg, endotoxin levels of pyrogen at 1.0 EU/mg, and bacterial DNA at 10 ng/mg. Pilot-scale fermentations and purifications were set up for the production of rhSCF that can be upscaled for industry.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Large‐scale purification and characterization of recombinant human stem cell factor in Escherichia coli

Chen

Cai

Zhang

et al. 2017

Biotech and App Biochem

View full text Add to dashboard Cite

show abstract

“…The critical problems are misfolding and aggregation, respectively. How to cope with these limitations in detail is out of the scope of this chapter and is discussed in specialized texts (60)(61)(62)(63)(64).…”

Section: The Use Of Microspheres In Protein Refoldingmentioning

confidence: 99%

“…Adsorption to ion exchangers is restricted to the use of the noncharged urea while hydrophobic interactions and biospecific affinity interactions can be applied in both denaturants. For a recent comprehensive review of the subject, the reader is referred to the text by Su et al (64).…”

Section: Solid-phase (On-column) Refoldingmentioning

confidence: 99%