{
  "sample_id": "ilknur_icke_2026-10-06_reteplase_pep_glutathione_balance_v1",
  "sample_type": "experimental",
  "reagent_list": {
    "lysate": 5000,
    "template_reteplase": 2000,
    "base_buffer": 2000,
    "magnesium_glutamate": 225,
    "ammonium_glutamate": 200,
    "amino_acid_mix_17": 400,
    "hepes_koh": 550,
    "pep_mono": 6000,
    "folinic_acid": 50,
    "coa": 100,
    "nad": 75,
    "putrescine": 50,
    "spermidine": 125,
    "oxidized_glutathione": 350,
    "reduced_glutathione": 625,
    "potassium_oxalate": 150,
    "dsbc_ecoli": 1000,
    "atp": 250,
    "ctp": 175,
    "gtp": 175,
    "utp": 175,
    "nuclease_free_water": 325
  },
  "metadata": {
    "ai_model": "Codex (OpenAI); exact model version not recorded",
    "protein_target": "Reteplase",
    "acknowledgements": "AI-assisted alternative based on course Reteplase PEP reference Reteplase_R2PC_041 and the simulated wet-lab-expert example. Prepared locally; no experiment or replacement submission is claimed.",
    "comments": "Baseline: Reteplase_R2PC_041. Change 170 mM GSSG from 475 to 350 nL and 100 mM GSH from 200 to 625 nL; water becomes 325 nL. Keep DsbC at 1,000 nL (5 uM) and all other additions fixed. Added GSSG/GSH become 2.975/3.125 mM; [GSH]+2[GSSG] remains 9.075 mM. Total 20,000 nL; all transfers are multiples of 25 nL.\n\nQuestion: does changing the added glutathione balance alter active Reteplase output at fixed DsbC? Compare with the unchanged reference in the same lysate and assay conditions, with replication. The published 312-well experimental control set held GSSG/GSH at 475/200 nL. No yield or actual redox potential is predicted. Higher activity would not by itself prove more correct disulfide pairing.\n\nAI-assisted preparation was checked against the course inventory. This design is a follow-up learning proposal and has not been experimentally tested."
  }
}