Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications

Byrne

Diamond and Related Materials

2015

Diamond-like carbon (DLC) coatings are useful for creating biocompatible surfaces for medical implants. DLC and silicon doped DLC have been synthesised using plasma enhanced chemical vapour deposition (PECVD). The effects of surface morphology on the interaction of human serum albumin (HSA) with doped and undoped DLC films have been investigated using a range of surface analysis techniques using Raman spectroscopy and atomic force microscopy (AFM). Raman spectra of doped DLC show that silicon doped DLC reduces the growth range of the ID/IG ratio, with a significant red-shift of the G peak position. Following exposure to protein, for undoped DLC the peaks at 1664 cm- 1 and around 1241 cm- 1 can be attributed to amide I and III, respectively, with an increase in the surface morphology of the surfaces giving some indication of the protein structure on the surfaces. Results indicate that HSA exhibit the majority of Î²-sheet during the adsorption on the surfaces. The results showed that the silicon incorporation DLC tends to increase of surface roughness and the adsorbed level of HSA is higher with higher levels of silicon doping of the DLC. Therefore, doping DLC may provide a method of controlling the adsorption of protein. Â© 2015 Elsevier B.V. All rights reserved

Section: Raman Spectroscopysupporting

confidence: 92%

Characteristic of silicon doped diamond like carbon thin films on surface properties and human serum albumin adsorption

Byrne

Diamond and Related Materials

2015

“…They are chemically and biologically stable and can be produced with ease and precise control. Other remarkable engineered antibody alternatives include single-chain variable fragments (ScFv) (111), camelid-derived heavy variablechain (VHH) antibodies (nanobodies) (112,113), single-chain antibodies expressed via yeast surface display (114), DARPins (115), and other artificial proteins such as adhirons (116). The advantages of these alternatives are that they are comparatively small, easily customized, and conveniently mass produced in bacterial systems, avoiding traditional antibody production in mammals or birds.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Biosensors for Whole-Cell Bacterial Detection

et al. 2014

SUMMARY Bacterial pathogens are important targets for detection and identification in medicine, food safety, public health, and security. Bacterial infection is a common cause of morbidity and mortality worldwide. In spite of the availability of antibiotics, these infections are often misdiagnosed or there is an unacceptable delay in diagnosis. Current methods of bacterial detection rely upon laboratory-based techniques such as cell culture, microscopic analysis, and biochemical assays. These procedures are time-consuming and costly and require specialist equipment and trained users. Portable stand-alone biosensors can facilitate rapid detection and diagnosis at the point of care. Biosensors will be particularly useful where a clear diagnosis informs treatment, in critical illness (e.g., meningitis) or to prevent further disease spread (e.g., in case of food-borne pathogens or sexually transmitted diseases). Detection of bacteria is also becoming increasingly important in antibioterrorism measures (e.g., anthrax detection). In this review, we discuss recent progress in the use of biosensors for the detection of whole bacterial cells for sensitive and earlier identification of bacteria without the need for sample processing. There is a particular focus on electrochemical biosensors, especially impedance-based systems, as these present key advantages in terms of ease of miniaturization, lack of reagents, sensitivity, and low cost.

“…Here we have screened our established Affimer phage library (Tiede et al, 2014) against a broad range of targets, including homologous protein family members, to isolate highly specific and renewable binding reagents that can be used both in vitro and in vivo. For broad applicability, and to remove the bottleneck in target protein production, we also tested our ability to generate reagents against small quantities of target protein from commercial sources.…”

Section: Introductionmentioning

confidence: 99%

Affimer proteins are versatile and renewable affinity reagents

Tiede

Bedford

Heseltine

et al. 2017

eLife

Self Cite

160

162

Molecular recognition reagents are key tools for understanding biological processes and are used universally by scientists to study protein expression, localisation and interactions. Antibodies remain the most widely used of such reagents and many show excellent performance, although some are poorly characterised or have stability or batch variability issues, supporting the use of alternative binding proteins as complementary reagents for many applications. Here we report on the use of Affimer proteins as research reagents. We selected 12 diverse molecular targets for Affimer selection to exemplify their use in common molecular and cellular applications including the (a) selection against various target molecules; (b) modulation of protein function in vitro and in vivo; (c) labelling of tumour antigens in mouse models; and (d) use in affinity fluorescence and super-resolution microscopy. This work shows that Affimer proteins, as is the case for other alternative binding scaffolds, represent complementary affinity reagents to antibodies for various molecular and cell biology applications.DOI: http://dx.doi.org/10.7554/eLife.24903.001