Stability Determination of Intact Humanin-G with Characterizations of Oxidation and Dimerization Patterns.

Ozgul M, Nesburn AB, Nasralla N, Katz B, Taylan E, Kuppermann BD, Kenney MC

Biomolecules11 Mar 2023Cited by 3 (Europe PMC)

PMID36979450PMCIDPMC10046509DOI10.3390/biom13030515PepBankPUB-36979450

Summary

Humanin is the first identified mitochondrial-derived peptide. Humanin-G (HNG) is a variant of Humanin that has significantly higher cytoprotective properties. Here, we describe the stability features of HNG in different conditions and characterize HNG degradation, oxidation, and dimerization patterns over short-term and long-term periods. HNG solutions were prepared in high-performance liquid chromatography (HPLC) water or MO formulation and stored at either 4 °C or 37 °C. Stored HNG samples were analyzed using HPLC and high-resolution mass spectrometry (HRMS). Using HPLC, full-length HNG peptides in HPLC water decreased significantly with time and higher temperature, while HNG in MO formulation remained stable up to 95% at 4 °C on day 28. HNG peptides in HPLC water, phosphate-buffered saline (PBS) and MO formulation were incubated at 37 °C and analyzed at day 1, day 7 and day 14 using HRMS. Concentrations of full-length HNG peptide in HPLC water and PBS declined over time with a corresponding appearance of new peaks that increased over time. These new peaks were identified to be singly oxidized HNG, doubly oxidized HNG, homodimerized HNG, singly oxidized homodimerized HNG, and doubly oxidized homodimerized HNG. Our results may help researchers improve the experimental design to further understand the critical role of HNG in human diseases.

CC BYText from Europe PMC. Highlighted sentences are the source of results.

Results from this study5

No result from this study comes from a test of the peptide.

Studies that did not test the peptide alone5

In this study, Humanin was a lab tool or a carrier, part of a mix, or the subject of a lab method. These results are not evidence about taking Humanin.

01LabNot a test of Humanin aloneIn vitro (cell-free)Other
In HPLC water, full-length Humanin-G decreased significantly with storage time and higher temperature.
In plain wordsIn lab tests, Humanin-G in water broke down more as storage time and temperature went up.
Peptide
Humanin
Outcome
DecreaseFull-length HNG in HPLC water over time and temperature
Study
n: not reported in source
Humanin-G in HPLC water, PBS or MO formulation
Outcome group
Formulation, delivery and stability · Pharmacology
Source sentences
Summary
Using HPLC, full-length HNG peptides in HPLC water decreased significantly with time and higher temperature
2 context sentences(design, population, supporting numbers)
Summary · context
HNG solutions were prepared in high-performance liquid chromatography (HPLC) water or MO formulation and stored at either 4 °C or 37 °C.
Summary · context
Stored HNG samples were analyzed using HPLC and high-resolution mass spectrometry (HRMS).
In this study, Humanin was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking Humanin.
Show in the study text Report a problemFND-3697945001
02LabNot a test of Humanin aloneIn vitro (cell-free)Other
In the MO formulation at 4 °C, full-length Humanin-G remained stable at up to 95% on day 28.
In plain wordsIn lab tests, Humanin-G stayed stable in the MO formulation at 4 °C for 28 days.
Peptide
Humanin
Outcome
No significant changeFull-length HNG in MO formulation at 4 °C on day 28
Study
n: not reported in source
Humanin-G in HPLC water, PBS or MO formulation
Outcome group
Formulation, delivery and stability · Pharmacology
Measurements
Full-length HNG remaining
≤ 95%
MO formulation, 4 °C, day 28
Source sentences
Summary
while HNG in MO formulation remained stable up to 95% at 4 °C on day 28.
2 context sentences(design, population, supporting numbers)
Summary · context
HNG solutions were prepared in high-performance liquid chromatography (HPLC) water or MO formulation and stored at either 4 °C or 37 °C.
Summary · context
Stored HNG samples were analyzed using HPLC and high-resolution mass spectrometry (HRMS).
In this study, Humanin was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking Humanin.
Show in the study text Report a problemFND-3697945002
03LabNot a test of Humanin aloneIn vitro (cell-free)Other
At 37 °C, full-length Humanin-G concentrations in HPLC water and PBS declined over time as new peaks appeared.
In plain wordsIn lab tests, Humanin-G in water and PBS declined over time at body temperature.
Peptide
Humanin
Outcome
DecreaseFull-length HNG in HPLC water and PBS at 37 °C
Study
n: not reported in source
Humanin-G in HPLC water, PBS or MO formulation
Outcome group
Formulation, delivery and stability · Pharmacology
Source sentences
Summary
HNG peptides in HPLC water, phosphate-buffered saline (PBS) and MO formulation were incubated at 37 °C and analyzed at day 1, day 7 and day 14 using HRMS. Concentrations of full-length HNG peptide in HPLC water and PBS declined over time with a corresponding appearance of new peaks that increased over time.
2 context sentences(design, population, supporting numbers)
Summary · context
HNG solutions were prepared in high-performance liquid chromatography (HPLC) water or MO formulation and stored at either 4 °C or 37 °C.
Summary · context
Stored HNG samples were analyzed using HPLC and high-resolution mass spectrometry (HRMS).
In this study, Humanin was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking Humanin.
Show in the study text Report a problemFND-3697945003
04LabNot a test of Humanin aloneIn vitro (cell-free)Other
Degradation of Humanin-G produced singly and doubly oxidized forms.
In plain wordsIn lab tests, Humanin-G broke down into oxidized forms.
Peptide
Humanin
Outcome
Not applicableHNG oxidation products
Study
n: not reported in source
Humanin-G in HPLC water, PBS or MO formulation
Outcome group
Formulation, delivery and stability · Pharmacology
Source sentences
Summary
These new peaks were identified to be singly oxidized HNG, doubly oxidized HNG, homodimerized HNG, singly oxidized homodimerized HNG, and doubly oxidized homodimerized HNG.
2 context sentences(design, population, supporting numbers)
Summary · context
HNG solutions were prepared in high-performance liquid chromatography (HPLC) water or MO formulation and stored at either 4 °C or 37 °C.
Summary · context
Stored HNG samples were analyzed using HPLC and high-resolution mass spectrometry (HRMS).
In this study, Humanin was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking Humanin.
Show in the study text Report a problemFND-3697945004
05LabNot a test of Humanin aloneIn vitro (cell-free)Other
Degradation of Humanin-G produced homodimerized forms, including singly and doubly oxidized homodimers.
In plain wordsIn lab tests, Humanin-G also formed pairs of linked peptide molecules.
Peptide
Humanin
Outcome
Not applicableHNG dimerization products
Study
n: not reported in source
Humanin-G in HPLC water, PBS or MO formulation
Outcome group
Formulation, delivery and stability · Pharmacology
Source sentences
Summary
These new peaks were identified to be singly oxidized HNG, doubly oxidized HNG, homodimerized HNG, singly oxidized homodimerized HNG, and doubly oxidized homodimerized HNG.
2 context sentences(design, population, supporting numbers)
Summary · context
HNG solutions were prepared in high-performance liquid chromatography (HPLC) water or MO formulation and stored at either 4 °C or 37 °C.
Summary · context
Stored HNG samples were analyzed using HPLC and high-resolution mass spectrometry (HRMS).
In this study, Humanin was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking Humanin.
Show in the study text Report a problemFND-3697945005

AI wrote and checked these results. AI can make mistakes, so check important results against the source.

Version history

PepBank keeps every version of the data on this study. Status changes from Retraction Watch, Europe PMC and Crossref are listed with their source.

Version 2Current
13 Oct 2026, 21:44 UTC
Study results updated
Version 1
13 Oct 2026, 16:31 UTC
First added · source update from Europe PMC

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