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How long does survodutide stay within specification at minus 80 °C once reconstituted?

Asked 23 Apr 2024Modified 2.0 years agoViewed 24k times
23

Conditions: survodutide · minus 80 °C.

I have done this once and I suspect I got away with it rather than got it right.

For context: I keep records of every batch, every lot number and every result, so an answer that requires me to track something is fine.

What does a defensible version of this look like in practice?

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JE
askedjuan_esquivel14k1623 Apr 2024
6Same situation here, so I will follow this one. – Dr_Ingrid_Baumgartner 7 months ago
7Is the material lyophilised or already in solution? Completely different answer. – Dr_Marek_Zielinski 9 months ago
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5 Answers

Accepted answer first, then by votes
166

Accepted answer

Whatever the refrigerated figure is, freezing does not simply extend it. 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. A frozen solution is not a slow solution: ice excludes solute, so the unfrozen fraction concentrates, the pH of the buffer shifts as one component crystallises first, and the damage happens during the transitions rather than during the hold. "Within specification" also needs a specification: purity, content, or both, and at what limit. Without that the question has no numerical answer at all.

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

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.

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.

Metal-catalysed oxidation of methionine is documented across peptide and protein formulations and is why chelators appear in some formulations.

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

edited 23 Jul 2024 by coldpack_88 — fixed an arithmetic slip in the third paragraph

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C8
answered · acceptedcoldpack_8850k3712 Jul 2024
8Adding for future readers: the domestic leg after delivery is the part you control. – nkem_obiora 2 months ago
Confirming that opening a cold vial in a humid room is a genuinely bad idea. – gradient_slope 4 months ago
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65

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

Hydrolysis cleaves the backbone, most readily at aspartate-proline and aspartate-glycine sequences, and is acid-catalysed. In a dry solid it barely proceeds at all.

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.

Deamidation via the succinimide intermediate is well characterised, with sequence-dependent rates highest for asparagine-glycine motifs.

A mass spectrum names the pathway. Plus one, plus sixteen, minus eighteen.

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LM
answeredleonid_marchuk19k2723 Jul 2024
49

The part that matters: aggregation is a physical process and is the one most often caused by handling rather than by time.

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.

Deamidation converts asparagine or glutamine to the corresponding acid via a succinimide intermediate, adding one dalton. It is base-catalysed, accelerates above neutral pH and is the dominant aqueous pathway for many peptides.

Cold, dry, dark, still. Those four words cover most of the mitigation.

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DV
answeredDr_Bram_Verhoeven84k24820 Jun 2024
7Two lots stored differently, reassayed at a year — the difference was smaller than I expected. – plate_count_9k 8 months ago
8The desiccant point is under-appreciated and costs nothing to act on. – lukas_sedlacek 2 days ago
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39

The short version: water enables most of it, oxygen enables oxidation, surfaces enable adsorption, and agitation enables aggregation.

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

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

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

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TH
answeredtyndall_haze38k381 Jul 2024
6Any published figure for how much a collapsed cake actually retains? – pieter_maas 4 months ago
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32

Answering this needs the physical state, since a dry powder is protected from most of these and a solution is protected from none.

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.

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

Sequence determines which pathways apply, so general statements are general.

At dilute concentrations, suspect adsorption before you suspect chemistry.

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JW
answeredj_wierzbicki69k14828 Apr 2024
The doubling-per-ten-degrees rule is the part I did not know and now use constantly. – anouk_desmet 4 months ago
2Small correction: it is the number of cycles rather than the freezer temperature that does the damage. – n_takahashi 6 months ago
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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.