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Aims: Women with previous gestational diabetes mellitus (GDM) have an increased risk for later development of type 2 diabetes. During pregnancy, GDM affects the cardio-metabolic protein profile; however, it is unknown how GDM affects the cardio-metabolic protein profile in the long term and if it is associated with type 2 diabetes after GDM. We hypothesise that the cardio-metabolic protein profile is affected long term and is associated with the development of type 2 diabetes after GDM., Materials and Methods: A case-control follow-up study based on the Odense GDM Follow-Up Study (OGFUS) cohort, which included women with previous GDM (n = 128) and matched controls without previous GDM (n = 70). Blood samples from a follow-up assessment 8-10 years after delivery were analysed using a 29-plex panel of apolipoproteins, transport and inflammation/immune proteins using multiple-reaction-monitoring mass spectrometry., Results: Apolipoprotein A-I, D and M were significantly lower in women with previous GDM compared to controls (all p < 0.001), while apolipoprotein L-I, H, vitamin D binding protein, CRP, vitronectin, transthyretin and complement factors 3 and B were significantly higher (p = 0.008, p < 0.001, p < 0.001, p < 0.001, p < 0.001, p = 0.005, respectively). All associations remained significant after adjustment for multiple testing except CRP, whereas apolipoprotein D, vitronectin and complement factors 3 and B were associated with the development of type 2 diabetes in women with previous GDM (p = 0.02, p = 0.001, p < 0.001, p = 0.004, respectively)., Conclusions: The cardio-metabolic protein profile 8-10 years after pregnancy is altered in women with previous GDM. Apolipoprotein D, vitronectin and complement factors 3 and B are candidate risk markers of type 2 diabetes after GDM pregnancy., (© 2025 The Author(s). Diabetes, Obesity and Metabolism published by John Wiley & Sons Ltd.)