Abstract
The impact of the microbiota on our health is rapidly gaining interest. While several bacteria have been associated with disease, and others being indicated as having a probiotic effect, the individual biomolecules behind these alterations are often not known. A major problem in the study of these factors in vivo is their low abundance in complex environments. We recently identified the first secreted bacterial antioxidant protein, RoxP, from the skin commensal Propionibacterium acnes, suggesting its relevance for maintaining the redox homeostasis on the skin. In order to study the effect, and prevalence, of RoxP in vivo, a capacitive biosensor with a recognition surface based on molecular imprinting was used to detect RoxP on skin in vivo. In vitro analyses demonstrated the ability to detect and quantify RoxP in a concentration range of 1 x 10−13 M to 1 x 10−8 M from human skin swabs; with a limit of detection of 2.5 x 10−19 M in buffer systems. Further, the biosensor was highly selective, not responding to any other secreted protein from P. acnes. Thus, it was possible to demonstrate the presence, and quantity, of RoxP on human skin. Therefore, the developed biosensor is a very promising tool for the detection of RoxP from clinical samples, offering a rapid, cost-effective and sensitive means of detecting low-abundant bacterial proteins in vivo in complex milieus.
Highlights
Propionibacterium acnes is one of the most common bacteria on human skin [1], and is involved in several skin disorders including acne vulgaris [2], and infections of prosthetic devices [3,4] due to biofilm formation on orthopedic implants [5]
Acrylamide, N-hydroxymethylacrylamide (NHMA), N-isopropylacrylamide (NIPAm), N,Nmethylenebisacrylamide (MBAAm), ammonium persulphate (APS), N,N,N’,N’-tetramethylethyldiamine (TEMED), Tris-HCl, Tween-20, Immunoglobulin A (IgA), Collagen, Bovine serum albumin (BSA), 3-aminopropyl-triethoxysilane (APTES), Glutaraldehyde (50%, w/v), Acryloylchloride, Triethylamine, 1-dodecanethiol, and Tyramine (99%) were all purchased from Sigma Aldrich
In comparison to a bare gold electrode (Fig 2A), the RoxP imprints on the RoxP-molecularly imprinted polymers (MIPs) electrode could be visualized as small non-distinctive cavities by scanning electron microscopy (SEM) (Fig 2B–2D)
Summary
Propionibacterium acnes is one of the most common bacteria on human skin [1], and is involved in several skin disorders including acne vulgaris [2], and infections of prosthetic devices [3,4] due to biofilm formation on orthopedic implants [5]. Mattiasson and Martin Hedstrom are associated with the company CapSenze, on whose instrument the analyses were performed, providing support in the form of instrument leasing, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. The other funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication
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