Sample preparation for glycoproteomics shipping checklist

Introduction

Pre‑analytical control is where glycan and protein integrity is won or lost. Hemolysis, proteolysis, or a single uncontrolled thaw can skew glycopeptide profiles, compress ratios, and derail downstream enrichment. This checklist codifies field‑tested steps that protect molecular integrity while meeting air-transport rules.

Scope: UN3373 Biological Substance, Category B under PI650, shipped with dry ice (UN1845) under PI954; research‑only. The focus is human plasma/serum, cell pellets, and snap‑frozen tissue for glycoproteomics.

Use this glycoproteomics shipping checklist to reduce compliance risk, avoid repacks and delays, and speed receiving approvals with clear acceptance criteria and complete documentation.

Risk matrix — common pre-analytical threats and mitigations

• Hemolysis → Impact: increased abundant hemoglobin peptides and oxidized glycans, masking low‑abundance glycopeptides. Mitigation: discard heavily hemolyzed samples; re-collect if possible; use strict venipuncture technique and immediate centrifugation.
• Proteolysis (post‑draw) → Impact: peptide cleavage alters glycopeptide ratios and site occupancy. Mitigation: add protease inhibitors at collection and freeze within ≤30 minutes; document timestamps.
• Uncontrolled thaw → Impact: glycan loss, altered glycopeptide distribution and ratio compression. Mitigation: aliquot ≤0.5 mL, avoid repeated freeze–thaw; quarantine and flag any thawed samples on receipt.

Key takeaways

• Use K2EDTA for plasma, add protease inhibitors, and freeze within ≤30 minutes when feasible; aliquot ≤0.5 mL to avoid repeat thaws.
• Pack to PI650: leakproof primary and secondary with absorbent; rigid outer; UN3373 diamond and "Biological Substance, Category B." According to the APHL Packing & Shipping Job Aid (2021), labels must be visible and not overlapped.
• Add dry ice per PI954 outside the secondary; vent outer; apply Class 9 label and mark "UN 1845 Dry ice, net kg." See the IATA DGR 66th Addendum (2025) documentation notes.
• For North America → Europe routes, maintain ≤–60°C (target –78.5°C) with a calibrated, NIST‑traceable logger; alert at ≥–50°C >30 minutes. Data logger battery exceptions are outlined in the IATA Lithium Battery Guidance.
• Include two AWB lines: "UN 3373, Biological Substance, Category B" and "UN 1845, Dry Ice, n × kg," as shown in Dartmouth EHS examples (2024).

Before you collect samples

Define study design and acceptance

• Confirm analytical workflow (e.g., HILIC or lectin enrichment, DIA vs. DDA) and library strategy; ensure sample prep supports the intended pipeline. For planning trade‑offs in DIA library strategy, see Creative Proteomics Glycoproteomics Service page.
• Pre‑define acceptance: matrices allowed, minimum volumes, tube types, label format, time‑to‑freeze target, aliquot sizes, and freeze–thaw limits (≤3 cycles). Record these in the protocol and manifest.

Align matrices, tubes, inhibitors

• Matrices/species in scope: human plasma (K2EDTA + protease inhibitor cocktail), human serum; cell pellets (human/rodent); snap‑frozen tissue.
• Tubes: 2.0 mL screw‑cap cryovials with O‑ring, low‑bind polypropylene; two external labels (human‑readable ID and 1D/2D barcode). Use solvent‑ and freeze‑tolerant labels.
• Inhibitors: For plasma, add protease inhibitors promptly per manufacturer guidance. Avoid introducing strong detergents or high chaotropes into material intended for submission.

For buffer composition and enrichment compatibility details (e.g., salts, detergents, HILIC vs. lectin enrichment), see Creative Proteomics' 4D Proteomics service overview for sample preparation considerations.

Time-to-freeze and aliquoting

• Operational target: ≤30 minutes from collection to initial freeze for biofluids/tissues; always record timestamps. This target aligns with minimizing cold ischemia windows discussed in NCI resources on snap‑freezing and best practices.
• Aliquoting: plasma/serum ≤0.5 mL per vial; cell pellets 1–5×10^6 cells per vial; tissue aliquots 20–50 mg. Avoid >3 freeze–thaw cycles. Store at ≤–80°C or LN₂ vapor until shipment. For practical QC advice on avoiding freeze–thaw pitfalls and variability in DIA, see common DIA pitfalls and fixes (Creative Proteomics glycoproteomics service ).

Cold-chain and packaging for your glycoproteomics shipping checklist

Triple packaging PI650

• Use leakproof primary receptacles inside a leakproof secondary with sufficient absorbent for liquids. Cushion and segregate multiple primaries so they don't contact each other. Place all within a rigid outer capable of a 1.2 m drop test.
• Mark the outer with the UN3373 diamond and the words Biological Substance, Category B adjacent to the mark. Keep label areas unobstructed and on a single face when practicable. For liquids, apply orientation arrows on two opposite sides. Summaries that mirror PI650 are compiled in the APHL job aid (2021).

Dry ice and LN2 PI954

• Place dry ice outside the secondary container and ensure the outer is vented to release CO₂. Affix the Class 9 miscellaneous hazard label and mark UN 1845 with the net dry ice weight (kg) on the outer. See the IATA DGR 66th Addendum (2025) for documentation and marking notes, and the FedEx Dry Ice DG Checklist (2025) for operator acceptance details.
• Typical planning mass for 30–48 h transit in an insulated shipper is ≥5–9 kg (adjust per container performance and validation). Ship early in the week to avoid weekend holds.

Lane validation and monitoring

• Scenario B (North America → Europe): maintain specimen environment ≤–60°C (target –78.5°C). Configure alerts at ≥–50°C sustained >30 minutes.
• Use a calibrated, NIST‑traceable temperature logger. Place it inside the outer near the secondary, not in direct contact with dry ice. Ensure any lithium batteries meet IATA data‑logger exceptions and UN38.3 testing, and use only the minimum number of devices needed.

Documentation and chain of custody

UN1845/UN3373 labels and marks

• Do not overlap labels. Keep the UN3373 diamond prominent and adjacent to Biological Substance, Category B. Apply the Class 9 label for dry ice on the same side as the dry ice marking and include UN 1845 with net dry ice kg. For overpacks, reproduce all required marks and add the word OVERPACK with letters ≥12 mm. These rules are reflected in IATA labels and markings resources.

Air waybill and invoices

• Include two lines on the AWB: "UN 3373, Biological Substance, Category B" and "UN 1845, Dry Ice, n × kg" (n = packages; kg = net dry ice per package). Add the responsible person's name and phone. Examples are shown in Dartmouth EHS shipping guidance (2024).

Records and calibration

• Retain the packing checklist, chain‑of‑custody with timestamps (collection, separation, initial freeze, packaging, pickup, receipt), and temperature logger calibration certificate (NIST‑traceable if claimed). Keep copies of operator training/authorization and any contracts for dangerous goods acceptance.

Pre-shipment consultation and manifest

Acceptance criteria

Disclosure: Creative Proteomics is our product.

Example intake line (≤30 words): ≥200 µL; 2.0 mL screw‑cap cryovials; dual labels & barcode; ≥5 kg dry ice/box; temp log + AWB docs; reject if thawed, damaged, mislabeled.

Sample manifest and barcoding

• Provide a manifest that maps each vial ID to its position in the shipper. Include human‑readable IDs and 1D/2D barcodes on vials and secondary pouches. List net dry ice kg, logger serial, start time, packer initials/date/time, and acceptance criteria checkboxes.

Pilot, excursions, escalation

• Pilot: For any new lane or container, run a mock or sacrificial shipment with a logger. Success criterion: no temperature ≥–50°C for >30 minutes. Archive the report for QA.
• Excursions: If alerts occur, quarantine on receipt, review the logger trace, packing method, dry ice mass, and route timings; document corrective and preventive actions.
• Escalation: Three tiers—Lab tech → PI/platform manager → carrier account/claims. Notify carrier within 2 hours of a confirmed incident and request timeline for investigation. Consider re‑icing plans only if the operator supports it and compliance is maintained.

Conclusion

• Do's: Freeze quickly (≤30 minutes when feasible), aliquot small volumes, use PI650 triple packaging, load sufficient dry ice with venting, mark UN3373/UN1845 clearly, and monitor with a calibrated logger. Keep a complete chain‑of‑custody and calibration records.
• Don'ts: Don't place dry ice inside the secondary, don't bury the logger in dry ice, don't overlap or tape over labels, don't ship late week without buffer dry ice.
• Next steps: Align your SOPs, manifests, and logger settings with your service provider and carrier before booking. If you need a quick review against the above acceptance checklist and lane thresholds, schedule a short coordination call.

References

1. Category B triple packaging summaries: the APHL Packing & Shipping Job Aid (2021).
2. Dry ice marking and net weight on the outer: IATA DGR 66th Addendum (2025) and FedEx Dry Ice DG Checklist (2025).
3. AWB phrase examples: Dartmouth EHS shipping guidance (2024).
4. Data logger and lithium battery exceptions: IATA Lithium Battery Guidance.

Author: Caimei Li, Senior Scientist, Creative Proteomics. Credentials: experienced in proteomics sample preparation and laboratory operations; public professional listings indicate active role in proteomics and metabolomics workflows. Version v1.1; Last updated: 2026-01-26.