Aim: Two approaches based on molecularly imprinted polymers-stir bar sorptive extraction (MIP-SBSE) and -magnetic solid-phase extraction (MIP-MSPE) have been used for extraction of carbamazepine (CBZ) from serum samples. Methodology: In MSPE and SBSE, development was achieved by employing a polycaprolactam coating. The Cecil® chromatographic system equipped with a UV-Vis detector was used for analytical determination […]
Aim: Novel bifunctional VEGF-A neutralizing therapies are being developed for the treatment of retinal vascular diseases such as age-related macular degeneration and diabetic retinopathy. In developing new therapeutic drugs, only small aqueous humor sample volumes are available for analyzing several parameters. Highly sensitive detection methods must be applied in analyzing VEGF-A levels in ocular fluids
Aim: The transitivity of osimertinib to cerebrospinal fluid (CSF) is of clinical concern. A quantitative LC–MS/MS method for the determination of osimertinib in human plasma and CSF was developed to evaluate its transitivity. Results: The calibration range was 40–1000 nM in plasma and 0.8–100 nM in CSF. Accuracy and precision were within 15%. Osimertinib in
Aim: A low-fouling and highly sensitive fluorescence immunoassay for protein detection in serum was proposed, and IgG was used as a model protein. Materials & methods: SH-PEG-NH2 serving as antifouling coating was conjugated with carboxyl Fe3O4 nanoparticles, and then, the thiol groups were conjugated with antibody via the covalent binding. IgG was captured through magnetic
Aim: Conjugated critical reagents are a pillar of ligand binding analysis. Although good practices for characterization have already been discussed, little is known about how assays are affected by specific conjugation parameters. Results: Here we developed, characterized and screened a toolset of bioconjugates that provided new insights about the optimization of conjugated critical reagent attributes.
In response to an earlier workshop covering the pros and cons of quantification below the LLOQ (BLQ) the author reviews the topics discussed from the bioanalytical standpoint. Important considerations for estimating concentrations below the LLOQ include: method signal-to-noise, baseline shape and condition, close lying interference peaks (especially for protein methods), matrix effect, adsorption and stability
In recent years, hybrid ligand-binding assays (LBAs)/LC–MS assays have been increasingly used for quantitation of protein biomarkers in biological matrices. However, unlike in LBAs where the importance of critical reagent screening and characterization is well understood and widely reported, benefits of well-characterized hybrid LC–MS assay reagents are frequently underestimated. Two groups of analyte-specific reagents, binding
Aim: Mucopolysaccharidosis type II (MPS II) is a lysosomal storage disorder caused by a deficiency of the iduronate-2-sulfatase enzyme leading to the accumulation of heparan sulfate (HS) and dermatan sulfate (DS) in organs and biological fluids. enzyme-replacement therapy is available for affected patients. Results/methodology: A 6-min UPLC–MS/MS method was developed/validated for HS and DS quantification
Aim: To develop a bioassay for estimation of sodium, potassium and creatinine in dried urine strips and comparing with their respective concentration in liquid urine samples. Materials & methods: Urine was collected on filter paper strips, dried at room temperature and, eluted for estimation of sodium, potassium by indirect ion selective electrode method and creatinine
Aim: Evolocumab is a human monoclonal antibody used in the treatment of cardiovascular diseases, which targeted proprotein convertase subtilisin kexin type 9. To accurately quantify free (including partially bound) and total evolocumab concentrations in serum, indirect and generic ELISA methods were developed and validated in rat serum. Results: Indirect ELISA was accurate and precise over