As a non-antibody scaffold, monobodies based on the fibronectin type III (FN3) domain overcome antibody size and complexity while maintaining analogous binding loops. However, antibodies and their derivatives remain the gold standard for the design of new therapeutics. In response, clinical-stage therapeutic proteins based on the FN3 domain are beginning to use native fibronectin function as a point of differentiation. The small and simple structure of monomeric monobodies confers increased tissue distribution and reduced half-life, whilst the absence of disulphide bonds improves stability in cytosolic environments. Where multi-specificity is challenging with an antibody format that is prone to mis-pairing between chains, multiple FN3 domains in the fibronectin assembly already interact with a large number of molecules. As such, multiple monobodies engineered for interaction with therapeutic targets are being combined in a similar beads-on-a-string assembly which improves both efficacy and pharmacokinetics. Furthermore, full length fibronectin is able to fold into multiple conformations as part of its natural function and a greater understanding of how mechanical forces allow for the transition between states will lead to advanced applications that truly differentiate the FN3 domain as a therapeutic scaffold.

No abstract

Compression therapy is the current evidence-based approach to manage venous leg ulcers (VLU); however, adherence is a major barrier to successful treatment. Combination approaches may relieve the burden of treatment by shortening the time to ulcer healing. This scoping review conducted by Australian researchers aimed to establish the evidence of effectiveness of various adjuvant methods on wound healing and recurrence. Randomized Controlled Trials (RCTs), and Systematic Reviews (SR) and Meta-Analyses (MA) on VLU management approaches published from January 2015 to December 2018 were included in this review. The articles included in the scoping review were grouped according to the management approaches, including (1) pharmaceutical interventions, (2) surgical interventions, (3) topical agents, (4) the use of devices, and (5) other, such as physiotherapy and psychological interventions. Results of this scoping review indicate that there is a limited high-quality evidence of effectiveness in most adjuvant therapies on wound healing and recurrence. Given the lowquality evidence observed in this scoping review for adjuvant treatments, the implication for practice is that current management guidelines be followed. Further rigorous studies have the potential to produce better quality evidence. Quality of evidence can be improved by ensuring large sample sizes of a single etiology wounds, standardizing reporting outcomes, and maintaining detailed and evidence-based protocols in physiological or psychological interventions.

Thyroid peroxidase (TPO) is a critical membrane-bound enzyme involved in the biosynthesis of multiple thyroid hormones, and is a major autoantigen in autoimmune thyroid diseases such as destructive (Hashimoto) thyroiditis. Here we report the biophysical and structural characterization of a novel TPO construct containing only the ectodomain of TPO and lacking the propeptide. The construct was enzymatically active and able to bind the patient-derived TR1.9 autoantibody. Analytical ultracentrifugation data suggest that TPO can exist as both a monomer and a dimer. Combined with negative stain electron microscopy and molecular dynamics simulations, these data show that the TR1.9 autoantibody preferentially binds the TPO monomer, revealing conformational changes that bring together previously disparate residues into a continuous epitope. In addition to providing plausible structural models of a TPO-autoantibody complex, this study provides validated TPO constructs that will facilitate further characterization, and advances our understanding of the structural, functional, and antigenic characteristics of TPO, an autoantigen implicated in some of the most common autoimmune diseases.

No abstract

Cobalt-mediated radical polymerizations (CMRPs) have been initiated by the radical decarboxylation of tetrachlorophthalimide activated esters. This allows for the controlled radical polymerization of activated monomers across a broad temperature range with a single cobalt species, with the incorporation of polymer end groups derived from simple carboxylic acid derivatives and termination with an organozinc reagent. This method has been applied to the synthesis of a polymer/graphene conjugate and a water-soluble protein/polymer conjugate, demonstrating the first examples of CMRP in graphene and protein conjugation.

25Thyroid peroxidase (TPO) is a critical membrane-bound enzyme involved in the biosynthesis of 26 multiple thyroid hormones, and is a major autoantigen in autoimmune thyroid diseases such as 27 Graves' disease and Hashimoto's thyroiditis. Here we report the biophysical and structural 28 characterisation of two novel TPO constructs containing only the ectodomain of TPO and lacking the 29 propeptide. Both constructs were enzymatically active and able to bind the patient-derived TR1.9 30 autoantibody. Analytical ultra-centrifugation data suggests that TPO can exist as both a monomer 31 and a dimer. Combined with negative stain electron microscopy and molecular dynamics 32 simulations, these data show that TR1.9 autoantibody preferentially binds the TPO monomer, 33 revealing conformational changes that bring together previously disparate residues into a 34 continuous epitope. In addition to providing plausible structural models of a TPO-autoantibody 35 complex, this study provides validated TPO constructs that will facilitate further characterization, 36 and advances our understanding of the structural, functional and antigenic characteristics of TPO, a 37 molecule behind some of the most common autoimmune diseases. 38 39 40 41 48 (Hashimoto's disease) and hyperthyroidism (Grave's disease) 2,3. AITDs are some of the most 49 common autoimmune diseases in the developed world, with Hashimoto's disease being a strong risk 50 factor for thyroid cancer 4. TPO is suspected to be involved in the pathogenesis of Hashimoto's 51 thyroiditis (prevalence 300-2980 cases per 100,000 in the Western world), leading to thyrocyte 52 destruction via CD8+ T-cell infiltration resulting in hypothyroidism 1,3,5. This incidence rate can be 53 compared to other autoimmune diseases -for example type 1 diabetes, which has an incidence in 54 the developed world of 310-570 cases per 100,000 patients -demonstrating the immense disease 55 burden caused by AITDs 5. The pathogeneses that underlies these autoimmune diseases is complex, 56 however the lack of any tertiary or quaternary structure of TPO complicates matters. The absence of 57 a structure in which to understand binding of anti-TPO antibodies, which are prevalent almost 58 ubiquitously (>95%) in cases of destructive thyroiditis, complicates the understanding of the 59 pathogenesis and nature of the disease. The precise mechanism by which these antibodies cause 60 damage is uncertain 2. Additionally, in some cases AITDs can occur without these autoantibodies 61 being present, and transplacental passage of anti-TPO antibodies does not necessarily cause thyroid 62 damage in the offspring 3,6,7. Despite this, transplacental passage of these autoantibodies can 63 potentially have cognitive effects on the child. In addition, antibodies to thyroglobulin (Tg) and 64 thyroid stimulating hormone receptor have been identified in both conditions, indicating that there 65 may be multiple antigens in AITD pathogenesis 2. 66TPO is a member of the animal heme peroxidase family, which also includes myelope...

As a non-antibody scaffold, monobodies based on the fibronectin type III (FN3) domain overcome antibody size and complexity while maintaining analogous binding loops. However, antibodies and their derivatives remain the gold standard for design of new therapeutics. In response, clinical therapeutic proteins based on the FN3 domain are beginning to use native fibronectin function as a point of differentiation. The small and simple structure of monomeric monobodies confers increased tissue distribution and reduced half-life, whilst the absence of disulphide bonds improves stability in cytosolic environments. Where multi-specificity is challenging with an antibody format that is prone to mis-pairing of chains, FN3 domains in the fibronectin assembly already interact with a large number of molecules. As such, multiple monobodies engineered for interaction with therapeutic targets are being combined in a similar beads-on-a-string assembly which improves both efficacy and pharmacokinetics. Furthermore, full length fibronectin is able to fold into multiple conformations as part of its natural function and a greater understanding of how mechanical forces allow for the transition between states will lead to advanced applications that truly differentiate the FN3 domain as a therapeutic scaffold.