PeptideStack
5.2kquestions
20kanswers
220users

What happens to dulaglutide after two weeks at minus 80 °C in solution?

Asked 24 Sept 2024Modified 20 months agoViewed 46k times
27

Setup, so nobody has to ask: dulaglutide · two weeks · minus 80 °C.

The empirical answer seems settled. The explanation does not.

If the honest answer is that nobody knows, I would rather hear that than a plausible story.

What is the causal chain, and where does it stop being established?

peptide-stability
peptide-stability

The chemistry of peptide degradation: deamidation, oxidation, hydrolysis, aggregation and fibrillation, and how temperature, pH, ionic strength,…

908 questions
storage
storage

Storage conditions and their evidence base: minus twenty degrees for powder, refrigerated for solution, protection from light, and what the…

701 questions
shelf-life
shelf-life

How long a preparation remains within specification: labelled expiry for a sealed lyophilised vial, beyond-use dating after reconstitution, and…

327 questions
dulaglutide
dulaglutide

A once-weekly GLP-1 receptor agonist built on an Fc fusion rather than fatty-acid acylation. Use this tag for questions about the fusion-protein…

200 questions
shareeditfollowflag
CC
askedcake_collapsed14k2724 Sept 2024
How many freeze-thaw cycles are we talking about? One and ten are different questions. – area_percent 6 months ago
Add the diluent — a preservative changes the in-use period entirely. – w_okoye 7 months ago
add a comment

3 Answers

Accepted answer first, then by votes
86

Accepted answer

two weeks is 14 days at a temperature where the chemistry all but stops and the physics does not. minus 80 °C is 85 kelvin below a refrigerator, and below the glass transition of a lyophilised cake the ten-degree rule of thumb stops applying at all — solid-state chemistry is not slow liquid chemistry, it is a different regime, and the failure modes that survive it are mechanical rather than chemical. In a frozen solution the solute is excluded from the growing ice, so the unfrozen fraction concentrates and the buffer's pH moves as one salt crystallises before the other. The damage is done at the transitions, and 14 days of stable hold between them contributes very little. Reconstituted material has no certificate; the one in the box describes the powder.

Start with the sequence, because which pathways are available depends on which residues are present.

Adsorption onto glass and plastic is significant at low concentrations — micrograms per millilitre — and negligible at milligrams per millilitre. It is the usual explanation for an apparent loss in a dilute preparation.

Aggregation is physical: peptides unfold at air-liquid interfaces and associate. Shaking maximises that interface, which is why swirling and shaking produce visibly different outcomes on the same vial.

Aggregation at air-liquid interfaces is established from surface-tension and particle-count studies and is the basis for anti-agitation handling guidance.

Nothing here is medical advice, and research-use compounds are not approved for human use.

Swirl, never shake. Aggregation is a handling problem more than a time problem.

shareimprove this answerflag
TV
answered · acceptedten_mg_vial31k1388 Nov 2024
Does the same reasoning apply to material already in solution, or is that a different curve? – p_mkhize 6 months ago
This should be linked from the help pages. – plate_count_9k 8 months ago
add a comment
Sponsored

Janoshik Analytical - Independent Third-Party Testing

HPLC purity, identity confirmation and quantified content on the vial you actually hold. Reports arrive with the chromatogram attached, not just a number.

Submit a sample
Sponsored — paired listing

GL Biochem (Shanghai) Ltd. - Direct Synthesis

Founded 1998. ISO 9001 and cGMP certified, 1,500+ staff and 200+ patents. The synthesis house behind a great many of the vials that get sent out for testing - batch-specific documentation with every order.

Visit GL Biochem
95

The relevant point is that a mass shift of plus one dalton is deamidation and plus sixteen is oxidation, so degradation is often visible in a mass spectrum if anyone looks.

Light exposure matters for tryptophan-containing sequences and for anything with a chromophore. Amber vials and a closed box are free mitigations.

To be exact about it, freeze-thaw cycling drives aggregation through concentration at the ice interface and pH shifts as buffer components crystallise out at different rates. Each cycle costs something.

Adsorption losses at low concentrations are quantified in formulation studies and are the reason carrier proteins are used in dilute preparations.

The caveat is that none of these pathways can be seen by looking at a vial, and a clear solution can be substantially degraded.

Sequence decides which pathways are even available. Check the residues.

shareimprove this answerflag
EL
answeredesben_lykke84k15816 Oct 2024
62

In practice, this is answerable from the chemistry rather than from anecdote, which is unusual and welcome.

Oxidation targets methionine, cysteine and tryptophan, adding sixteen daltons per oxygen. It is catalysed by trace metals and promoted by dissolved oxygen and by light.

A mass spectrum resolves most of this: minus eighteen is dehydration or succinimide, plus one is deamidation, plus sixteen is oxidation, and an unchanged mass with a shifted retention time is an isomer.

At dilute concentrations, suspect adsorption before you suspect chemistry.

edited 25 Nov 2024 by shear_at_the_front — clarified the distinction between purity and content

shareimprove this answerflag
SF
answeredshear_at_the_front17k2727 Oct 2024
3Small correction: it is the number of cycles rather than the freezer temperature that does the damage. – Dr_Yusuf_Adeyemi 8 months ago
add a comment

Your answer

Ask PeptideStack is a static archive. Posting is closed, but the norms are worth stating: answer the question that was asked, show your working, cite the trial or the certificate, and say plainly where the evidence runs out.

Not medical advice. Research-use-only compounds are not approved for human use.