Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Production of Pharmaceuticals01:30

Production of Pharmaceuticals

26
Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under...
26
Protein Folding Quality Check in the RER01:29

Protein Folding Quality Check in the RER

5.5K
ER is the primary site for the maturation and folding of soluble and transmembrane secretory proteins. The calnexin cycle is a specific chaperone system that folds and assesses the confirmation of N-glycosylated proteins before they can exit the ER lumen. The primary players of this quality check pipeline are the lectins, ER-resident chaperones, and a glucosyl transferase enzyme. In case the calnexin system in the lumen fails to salvage a misfolded protein, it is transported to the cytoplasm...
5.5K
Insulin: Biosynthesis, Chemistry, and Preparation01:25

Insulin: Biosynthesis, Chemistry, and Preparation

1.9K
The endoplasmic reticulum (ER) of pancreatic β-cells synthesizes preproinsulin, which consists of a signal peptide, A and B chains, and a C-peptide. Preproinsulin is then cleaved and folded into proinsulin, which translocates to the Golgi apparatus for sorting and packaging into secretory granules. In these granules, enzymatic clipping generates insulin and C-peptide.
Damage or functional impairment of β-cells inhibits insulin production, leading to diabetes. Diabetes treatment...
1.9K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Metabolic and inflammatory burden modifies muscle cardiovascular association across aging cohorts within an intrinsic capacity framework.

iScience·2026
Same author

Comprehensive pan-cancer analysis of FSCN1 as a marker for prognosis and immunity.

Discover oncology·2026
Same author

A cross-scale multimodal framework identifies clinically actionable immunotherapy biomarkers in melanoma through bulk to single-cell and spatial transcriptomics integration.

Human genomics·2026
Same author

Effect of <i>Polygonatum cyrtonema</i> Flour Addition on the Rheological Properties, Gluten Structure Characteristics of the Dough and the In Vitro Digestibility of Steamed Bread.

Foods (Basel, Switzerland)·2025
Same author

Separation Principles and Strategies for an Oil-Water Separation Membrane with Special Wettability.

Membranes·2025
Same author

Carbonized Dual-Layer Balsa Wood Membrane for Efficient Oil-Water Separation in Kitchen Applications.

Membranes·2025

Related Experiment Video

Updated: Mar 26, 2026

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
14:25

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Published on: December 12, 2017

19.1K

Improving the refolding efficiency for proinsulin aspart inclusion body with optimized buffer compositions.

Ying Chen1, Qi Wang1, Chun Zhang2

  • 1National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.

Protein Expression and Purification
|January 31, 2016
PubMed
Summary

Recovering native proinsulin conformation is key for efficient insulin manufacturing. Arginine and specific oxidants enhance refolding yields, improving disulfide bond formation and enabling high-purity insulin production.

Keywords:
AdditiveAggregateDiselenideInclusion bodiesOxidative refoldingProinsulin

More Related Videos

Real-time Analyses of Retinol Transport by the Membrane Receptor of Plasma Retinol Binding Protein
14:32

Real-time Analyses of Retinol Transport by the Membrane Receptor of Plasma Retinol Binding Protein

Published on: January 28, 2013

14.2K
Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous &#946;2-Microglobulin
11:17

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin

Published on: March 10, 2021

6.9K

Related Experiment Videos

Last Updated: Mar 26, 2026

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
14:25

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Published on: December 12, 2017

19.1K
Real-time Analyses of Retinol Transport by the Membrane Receptor of Plasma Retinol Binding Protein
14:32

Real-time Analyses of Retinol Transport by the Membrane Receptor of Plasma Retinol Binding Protein

Published on: January 28, 2013

14.2K
Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous &#946;2-Microglobulin
11:17

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin

Published on: March 10, 2021

6.9K

Area of Science:

  • Biochemistry
  • Protein Chemistry
  • Industrial Biotechnology

Background:

  • Efficient insulin manufacturing relies on native proinsulin conformation recovery from inclusion bodies.
  • Disulfide-linked oligomers and isomers are identified by-products during proinsulin refolding.

Purpose of the Study:

  • To investigate the distinct mechanisms of arginine and urea in proinsulin refolding.
  • To identify optimal conditions for enhancing native disulfide bond formation and overall refolding yield.
  • To evaluate the feasibility of industrial-scale proinsulin purification.

Main Methods:

  • Comparative analysis of arginine and urea effects on proinsulin refolding.
  • Identification and quantification of refolding by-products using multiple analytical techniques.
  • Optimization of refolding conditions using oxidants like selenocystamine.
  • One-step anionic exchange chromatography for purification.

Main Results:

  • Arginine increased refolding yield to 50% by reducing oligomers, while urea at higher concentrations increased disulfide-isomerized monomers, decreasing yield.
  • An oxidant, selenocystamine, combined with arginine, boosted yield to 80%.
  • Refolding with 2 M urea and reduced protein concentration (0.3 mg/mL) yielded high purity (95%) proinsulin via single-step chromatography.

Conclusions:

  • Arginine and urea exhibit different mechanisms and efficacies in proinsulin refolding.
  • Oxidants are crucial for native disulfide bond formation, not redox pairs.
  • Optimized refolding and purification strategies offer potential for improved industrial insulin production.