Towards a generic prototyping approach for therapeutically-relevant peptides and proteins in a cell-free translation system.

Advances in peptide and protein therapeutics increased the need for rapid and cost-effective polypeptide prototyping. While in vitro translation systems are well suited for fast and multiplexed polypeptide prototyping, they suffer from misfolding, aggregation and disulfide-bond scrambling of the translated products. Here we propose that efficient folding of in vitro produced disulfide-rich peptides and proteins can be achieved if performed in an aggregation-free and thermodynamically controlled folding environment. To this end, we modify an E. coli-based in vitro translation system to allow co-translational capture of translated products by affinity matrix. This process reduces protein aggregation and enables productive oxidative folding and recycling of misfolded states under thermodynamic control. In this study we show that the developed approach is likely to be generally applicable for prototyping of a wide variety of disulfide-constrained peptides, macrocyclic peptides with non-native bonds and antibody fragments in amounts sufficient for interaction analysis and biological activity assessment.

Clinical supervision: what do we know and what do we need to know? A review and commentary.

AIMS: This study is addressed to nurses but the issues are of equal concern to both midwives and health visitors. Clinical supervision ideally both challenges nurses as well as help their practice. There is need to identify critical elements that help professional practice and understand more clearly the changing nature of supervisory relationships. BACKGROUND: Clinical supervision in nursing is over a decade old in the UK and yet emerging nursing literature suggests that many ideas remain unfamiliar to nursing practice. The resistance shown by nurse towards clinical supervising remains perplexing. Moreover, ideas concerning clinical supervision have been applied without a substantive evidence base. METHODS: The discussion draws on varied ideas concerning supervision, including those outside of nursing, to ask what do we know and still need to know about clinical supervision. This study suggests that, a single approach to clinical supervision could be unhelpful to nursing. FINDINGS AND CONCLUSION: Nursing knowledge concerning many aspects of clinical supervision is increasing because of research. Much of the literature suggests that clinical supervision is scholarly activity requiring much the same attention to relationships as the therapeutic activities it supports. This discussion concludes with the idea that clinical supervision might work at its best as a quiet activity allowing nurses to think about nursing work in ways that suit individual learning styles.