A sensitive and selective approach for detection of IL 1α cancer biomarker using disposable ITO electrode modified with epoxy-substituted polythiophene polymer.

A highly sensitive and label-free electrochemical immunosensor for sensitive detection of interleukin 1α cancer biomarker was described by using epoxy-substituted polythiophene polymer modified disposable indium tin oxide electrode. This conjugated polymer was synthesized to fabricate this immunosensor for the first time and it offered a good biosensing platform for anti-IL 1α antibody immobilization due to epoxy groups present on the side groups of the polymer. Furthermore, the epoxy-substituted polythiophene polymer coated indium tin oxide electrode was used for the determination of IL 1α antigen for the first time. Electrode fabrication stages were characterized by using electrochemical (electrochemical impedance spectroscopy and cyclic voltammetry) and morphological (scanning electron microscopy and atomic force microscopy) techniques. Under optimum experimental conditions, impedimetric responses of the immunosensor increased with the increasing of IL 1α antigen concentration, and the proposed immunosensor displayed a wide linear detection range from 0.01 pg/mL to 5.5 pg/mL with a detection limit of 3.4 fg/mL. The proposed immunosensor exhibited outstanding performance including excellent reproducibility, good repeatability, high selectivity and long storage stability. With the advantages of simple operation, low-cost fabrication and label-free format, the suggested immunosensor was expected to have potential applications for IL 1α cancer biomarker detection.

Minimally-invasive Sampling of Interleukin-1α and Interleukin-1 Receptor Antagonist from the Skin: A Systematic Review of In vivo Studies in Humans.

Interleukin-1α (IL-1α) and its receptor antagonist IL-1RA play a pivotal role in skin homeostasis and disease. Although the use of biopsies to sample these cytokines from human skin is widely employed in dermatological practice, knowledge about less invasive, in vivo sampling methods is scarce. The aim of this study was to provide an overview of such methods by systematically reviewing studies in Medline, EMBASE, Web of Science and Cochrane Library using combinations of the terms "IL-1α", IL-1RA", "skin", "human", including all possible synonyms. Quality was assessed using the STrengthening the Reporting of OBservational studies in Epidemiology (STROBE) checklist. The search, performed on 14 October 2016, revealed 10 different sampling methods, with varying degrees of invasiveness and wide application spectrum, including assessment of both normal and diseased skin, from several body sites. The possibility to sample quantifiable amounts of cytokines from human skin with no or minimal discomfort holds promise for linking clinical outcomes to molecular profiles of skin inflammation.

[Preparation and application of polyclonal antibody against mouse IL-1α].

AIM: To construct a recombinant plasmid encoding mouse IL-1α (mIL-1α), express and purify mIL-1α protein, and prepare its polyclonal antibody. METHODS: The cDNAs were obtained from the spleen cells of BALB/c mice and the full length of mIL-1α gene was amplified by RT-PCR. Then the mIL-1α gene was inserted into a prokaryotic expression vector pET32a(+) and the resulting recombinant plasmid was transformed into E.coli BL21(DE3). After auto-induction, the mIL-1α protein was expressed and purified by electro-elution. An anti-mIL-1α polyclonal antibody was raised in New Zealand rabbits after immunization with the purified mIL-1α and the titer was determined by ELISA. The specificity of the polyclonal antibody was identified by Western blot and flow cytometry. RESULTS: The recombinant prokaryotic expression vector pET32a(+)-IL-1α was successfully constructed, and the mIL-1α protein was expressed and purified. ELISA showed the titer of the anti-mIL-1α serum was 1:25 600. Western blot and flow cytometry demonstrated the high specificity of the polyclonal antibody to IL-1α. CONCLUSION: The rabbit anti-mIL-1α polyclonal antibody with high titer and specificity has been prepared after immunization with the purified mIL-1α protein, facilitating further functional studies of IL-1α.