Identification of Lysine Misincorporation at Asparagine Position in Recombinant Insulin Analogs Produced in E. coli. View Full Text


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Article Info

DATE

2019-06

AUTHORS

Dorota Stadnik, Anna Bierczyńska-Krzysik, Joanna Zielińska, Jarosław Antosik, Piotr Borowicz, Elżbieta Bednarek, Wojciech Bocian, Jerzy Sitkowski, Lech Kozerski

ABSTRACT

PURPOSE: Identification of human insulin analogs' impurity with a mass shift +14 Da in comparison to a parent protein. METHODS: The protein sequence variant was detected and identified with the application of peptide mapping, liquid chromatography, tandem mass spectrometric analysis, nuclear magnetic resonance spectroscopy (NMR) and Edman sequencing. RESULTS: The misincorporated lysine (Lys) at asparagine (Asn) position A21 was detected in recombinant human insulin and its analogs. CONCLUSIONS: Although there are three asparagine residues in the insulin derivative, the misincorporation of lysine occurred only at position A21. The process involves G/U or A/U wobble base pairing. More... »

PAGES

79

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s11095-019-2601-z

DOI

http://dx.doi.org/10.1007/s11095-019-2601-z

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1113185257

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30949841


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43 schema:description PURPOSE: Identification of human insulin analogs' impurity with a mass shift +14 Da in comparison to a parent protein. METHODS: The protein sequence variant was detected and identified with the application of peptide mapping, liquid chromatography, tandem mass spectrometric analysis, nuclear magnetic resonance spectroscopy (NMR) and Edman sequencing. RESULTS: The misincorporated lysine (Lys) at asparagine (Asn) position A21 was detected in recombinant human insulin and its analogs. CONCLUSIONS: Although there are three asparagine residues in the insulin derivative, the misincorporation of lysine occurred only at position A21. The process involves G/U or A/U wobble base pairing.
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