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Peptides, as targeted therapeutics for metabolic disorders and cancer, have profoundly reshaped our modern pharmaceutical industry. Understanding the pharmacokinetic behaviour and pharmacological/toxicological effects of these drugs critically depends on determining the unbound fraction (fu) in plasma. However, inherent challenges such as stability and non-specific binding issues make obtaining reliable fu values particularly difficult for peptides using traditional plasma protein binding (PPB) experiments, such as equilibrium dialysis, ultracentrifugation, or ultrafiltration. To address these issues, this study developed a biomimetic chromatography method to evaluate human plasma protein binding.
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