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Insulin: Biosynthesis, Chemistry, and Preparation

Insulin: Biosynthesis, Chemistry, and Preparation

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...

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Epigenetic Conversion as a Safe and Simple Method to Obtain Insulin-secreting Cells from Adult Skin Fibroblasts

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Development of a human insulin certified reference material with SI-traceable purity.

Xianxia Wang¹, Liqing Wu², Yanjie Huang³

¹Beijing University of Chemical Technology, No. 15 North Third Ring Road, Chaoyang District, Beijing, 100029, China.

Analytical and Bioanalytical Chemistry

|February 24, 2022

Summary

A new human insulin (hINS) certified reference material (CRM) was developed for accurate quantification. This CRM ensures quality control in biopharmaceutical production and clinical diagnostics.

Keywords:

Human insulin certified reference material Isotope dilution mass spectrometry Liquid chromatography-circular dichroism Mass balance

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Area of Science:

Metrology
Biochemistry
Analytical Chemistry

Background:

Accurate quantification of human insulin (hINS) is crucial for biopharmaceutical production and clinical diagnostics.
Certified Reference Materials (CRMs) are essential for ensuring the reliability and comparability of analytical measurements.
The National Institute of Metrology (NIM) has developed a novel hINS CRM to meet these needs.

Purpose of the Study:

To develop and characterize a human insulin certified reference material (CRM).
To establish a reliable primary calibrator for insulin quantification.
To support quality control in biopharmaceutical manufacturing and clinical settings.

Main Methods:

Candidate material identification using de novo sequencing via mass spectrometry and Edman degradation.
Determination of insulin-related impurities, aggregation, moisture, volatile organic compounds (VOCs), anions, and ignition residues.
Certified value assessment using mass balance (MB) and amino acid analysis-based isotope dilution mass spectrometry (AAA-IDMS).
Validation of the certified value using liquid chromatography-circular dichroism spectroscopy (LC-CD) and quantitative nuclear magnetic resonance (qNMR).
Homogeneity and stability examinations under various storage conditions.

Main Results:

The hINS CRM was successfully developed and characterized.
The certified value was determined to be (0.857 ± 0.024) g/g.
Validation methods (LC-CD, qNMR) showed good agreement with the certified value.
The material demonstrated homogeneity.
The CRM exhibited stability for at least 12 months at -70°C and 7 days at elevated temperatures (4°C, 25°C, 40°C).

Conclusions:

The developed hINS CRM is a reliable primary calibrator.
This CRM will enhance quality control in biopharmaceutical production.
It will also support accurate diagnostics in clinical settings.
The characterization and validation confirm its suitability for intended applications.