Tailored pretreatment of serum samples and biomarker extraction afforded by ionic liquids as constituents of aqueous biphasic systems

resumo

Prostate cancer serum biomarkers, such as the U.S. Food and Drug Administration (FDA)-approved prostate specific antigen (PSA), are valuable markers of diagnosis, response to treatment and disease progression. Despite the expected benefits of using PSA to better manage the mortality and morbidity associated to prostate cancer, sample complexity and interference within analysis make its quantification in serum a high-cost and strenuous assignment. Here, we make use of the simple preparation, water-rich environment, and selectivity of aqueous biphasic systems (ABS) to develop alternative serum pretreatment strategies. A set of ammonium- and phosphonium-based ionic liquids (ILs) were used as either main phase-forming agents in IL-salt ABS or as adjuvants in polymer-salt ABS, allowing to explore the IL designer solvent nature and versatile role in ABS formation and performance. Serum pretreatment efficiency was evaluated by the capacity of each ABS to simultaneously deplete high abundance serum proteins (immunoglobulin G, IgG, and human serum albumin, HSA) at the systems' interphase (thus, generating a three-phase partitioning system - ABS-TPP) and extract PSA to a single ABS phase. Regardless of the ABS typology under appraisal (IL-salt or polymer-salt-IL), the proper design of the IL structure is critical for both the ability to deplete high abundance serum proteins and extract PSA. The results demonstrate that, in addition to the proper choice of the ABS typology, a careful adjustment of the IL hydrophobicity is key to achieve the simultaneous depletion of high-abundance proteins and complete PSA extraction in a single step, creating new approaches for PSA analysis and its application in diagnosis/prognosis.

palavras-chave

PROSTATE-SPECIFIC ANTIGEN; PHASE-DIAGRAMS; CANCER; INTERFERENCE; ASSAY; IMMUNOASSAYS; PURIFICATION; PROGRESSION; PROTEINS; PLATFORM

categoria

Engineering

autores

Rosa, ME; Mendes, MSM; Carmo, E; Conde, JP; Coutinho, JAP; Freire, MG; Silva, FAE

nossos autores

agradecimentos

This work was developed within the scope of the project CICECO- Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020, financed by national funds through the FCT/MEC (PIDDAC) , and the Research Unit INESC MN (UID/05367/2020) through BASE and PROGRAMATICO financing. This work was developed within the project PTDC/EMD-TLM/3253/2020 (ILSurvive) , funded by national funds (OE) , through FCT/MCTES. M.E.R. and M.S.M.M. and acknowledge FCT for the doctoral grants SFRH/BD/136995/2018 and 2022.11229.BD, respectively. F.A.eS. acknowledges FCT for the researcher contract CEECIND/03076/2018 under the Scientific Employment Stimulus - Individual Call 2018.

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