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A new method for the selective determination of laminin-5 concentration using a biosensor and surface plasmon resonance imaging (SPRI) technique is presented. A biosensor based on the specific interaction of laminin-5 with rabbit polyclonal antibody was constructed. The analytically useful dynamic response range of the biosensor is between 0.014 and 0.1 ng mL−1. The detection limit is 4 pg mL−1. The potential influence of interferences on the SPRI signal was investigated, and the high selectivity of the biosensor was confirmed. In order to demonstrate the potential application of the biosensor, laminin-5 concentration in blood plasma was determined. The results were compared with the laminin-5 concentration obtained by the commercial enzyme-linked immunosorbent assay (ELISA) kit. A comparison of results from healthy donors obtained by SPRI measurement and ELISA indicates that they are close and shows good agreement with the data reported in the literature. The plasma samples of bladder cancer patients gave higher concentration measured with specific biosensor than by ELISA assay. The study shows the clear difference in concentration of laminin-5 in healthy humans and patients with bladder cancer. Extensive clinical studies using the newly developed method can result in an increase in the use of laminin-5 as a potential cancer marker.Electronic supplementary materialThe online version of this article (doi:10.1007/s00216-016-9621-x) contains supplementary material, which is available to authorized users.

Aromatase (ARO) is an enzyme with potential diagnostic significance. Aberrant expression of aromatase in tissues is associated with a number of pathological conditions, including tumor of the breast, ovary, testes, liver, adrenal cortex and uterus, as well as endometriosis.Two methods for the highly selective determination of ARO concentration in human tissues by using of two different biosensors co-operating with the surface plasmon resonance imaging technique (SPRI) have been developed. One of the developed biosensors contains immobilised rabbit polyclonal antibody specific for aromatase (Y-ARO), while the other contains immobilized ARO inhibitor-exemestane (E-ARO). Both biosensors specifically bound ARO from analyzed samples. The analytically useful dynamic response range of both biosensors is between 0.3 and 5.0 ng mL−1. The detection limit (3S.D.) of both biosensors is 90 pg mL−1. Standard deviation of both biosensors is 1%. Recoveries of ARO spikes are between 97 and 108% for both biosensors under model conditions and for real samples. Albumin and alkaline phosphatase are tolerated for both biosensors up to 10,000 fold excess.

The 20S proteasome is a multicatalytic enzyme complex responsible for intracellular protein degradation in mammalian cells. Its antigen level or enzymatic activity in blood plasma are potentially useful markers for various malignant and nonmalignant diseases. We have developed a method for highly selective determination of the 20S proteasome using a Surface Plasmon Resonance Imaging (SPRI) technique. It is based on the highly selective interaction between the proteasome’s catalytic β5 subunit and immobilized inhibitors (the synthetic peptide PSI and epoxomicin). Inhibitor concentration and pH were optimized. Analytical responses, linear ranges, accuracy, precision and interferences were investigated. Biosensors based on either PSI and epoxomicin were found to be suitable for quantitative determination of the proteasome, with a precision of ±10% for each, and recoveries of 102% and 113%, respectively, and with little interference by albumin, trypsin, chymotrypsin, cathepsin B and papain. The proteasome also was determined in plasma of healthy subjects and of patients suffering from acute leukemia. Both biosensors gave comparable results (2860 ng·mL-1 on average for control, and 42300 ng·mL-1 on average for leukemia patients).FigureThe synthetic peptide aldehyde Z-Ile-Glu(OBut)-Ala-Leu-H (PSI) and a microbial α’,β’ epoxyketone peptide epoxomicin was used to develop SPRI biosensor for the highly selective determination of the 20S proteasome concentration, and to evaluate the sensor applicability for the determination of 20S proteasome in human blood plasma.