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The aim is to explore challenges related to the integration between product development and production in product introduction and, given these challenges, to analyse the learning potential of boundary crossing in the context of product introduction. The paper draws on evidence from a Swedish manufacturing company. The theoretical framework is based on a boundary-crossing perspective, which in turn is framed by a workplace learning perspective. Data were collected through interviews with 19 employees from the product development department and 21 employees from the production plant, and 8 focus-group interviews. Within the company, there were many challenges related to product introduction, but the findings also show these challenges can provide learning opportunities by enabling the boundaries to be crossed between the product development department and the production plant. Several forms of intrapersonal or interpersonal boundary crossing were identified. Individuals acted as brokers, and prototypes, pre-series, DfA analysis and a crossfunctional team served as boundary objects and encounters. Nothing in our study indicates that the boundary crossing identified on the intrapersonal and interpersonal levels created learning potentials on the organisational level in the company. The conclusion is that it is necessary to consider the learning potential made available by boundary crossing in order to support learning, and thereby improve the integration between product development and production in product introduction. By seeing and using prototypes and pre-series production as learning opportunities you can create a better preparedness and provide collective access to knowledge required for successful product introduction.

Despite that a decade has passed since the Industry 4.0-era began, there is still limited knowledge about the implications for the wooden single-family house industry. The purpose of this paper is to identify components of smart manufacturing that captures what companies in the wooden single-family house industry need to consider meeting the expected and future demands related to the fourth industrial revolution. The results presented is based on two case studies, revealing seven components. The industry is in the beginning of its implementation phase and is mainly striving for connectivity through integration of ICTs. [ABSTRACT FROM AUTHOR]

Abstract: Today''s global competitiveness urges SMMEs to pay attention to their MS process. The purpose of this case study at a Swedish SMME, mainly conducted through interviews with production-related staff: staff with direct connection to everyday production work, is to explore their perception of the MS content. The study shows that communication is the main obstacle for production-related staff''s perception of the MS. Their perception is diverse and based on a multitude of factors, such as employment period, organizational belonging, and the employees’ own interest. Several problem areas are identified and need to be investigated further. [Copyright &y& Elsevier]

Biacore systems (Biacore AB) enable label-free detection of molecular interactions in real time using surface plasmon resonance detection. Epitope mapping of antibodies is usually performed in a pairwise fashion where one antibody is used to capture the antigen from solution and the binding of a secondary antibody is monitored. In contrast to alternative approaches, the method allows for mapping of large matrices of antibody pairs without the need for cumbersome labeling steps, and the real-time analysis enables a better ranking of complex stability when compared with end-point assays.

Advances in molecular evolution strategies have made it possible to identify antibodies with exquisite specificities and also to fine-tune their biophysical properties for practically any specified application. Depending on the desired function, antibody/antigen interactions can be long-lived or short-lived and, therefore, particular attention is needed when seeking to identify antibodies with specific reaction-rate and affinity properties. Surface plasmon resonance (SPR) biosensors routinely generate sensitive and reliable kinetic data from antibody/antigen interactions for both therapeutic and diagnostic applications. However, many kinetic-based screening assays require rigorous sample preparation and purification prior to analysis. To ameliorate this problem, we developed a rapid and reliable assay for characterising recombinant scFv antibody fragments, directly from crude bacterial lysates. Ninety-six scFv antibodies derived from chickens immunised with C-reactive protein (CRP) were selected by phage display and evaluated using the Biacore A100 protein interaction array system. Antibodies were captured from crude bacterial extracts on the sensor chip surface and ranked based on the percentage of the complex left (% left) after dissociation in buffer. Kinetic rate constants (k(a) and k(d)) and affinity (K(D)) data were obtained for six clones that bound monomeric CRP across a broad affinity range (2.54 x 10(-8) to 3.53 x 10(-10) M). Using this assay format the A100 biosensor yielded high quality kinetic data, permitting the screening of nearly 400 antibody clones per day.

Label-free optical biosensor technology has become a standard tool for characterizing monoclonal antibodies for therapeutic and diagnostic applications. The availability of high-quality binding data at an early stage greatly improves the ability to select antibodies for further development. This article shows how Biacore A100, a protein interaction array system, is capable of providing high-quality data with increased throughput. In a 12-h automated run, we analyzed 386 crude hybridoma samples to identify those with the desired kinetic profiles. Selected antibodies were further characterized by higher resolution analysis, and binding interactions were studied under a range of buffer conditions. We demonstrate how this new parallel processing system significantly expands the throughput of protein interaction analysis while maintaining data quality.

The proto-oncogene c-myc governs the expression of a number of genes targeting cell growth and apoptosis, and its expression levels are distorted in many cancer forms. The current investigation presents an analysis by proteolysis, circular dichroism, fluorescence and Biacore of the folding and ligand-binding properties of the N-terminal transactivation domain (TAD) in the c-Myc protein. A c-Myc sub-region comprising residues 1-167 (Myc1-167) has been investigated that includes the unstructured c-Myc transactivation domain (TAD, residues 1-143) together with a C-terminal segment, which appears to promote increased folding. Myc1-167 is partly helical, binds both to the target proteins Myc modulator-1 (MM-1) and TATA box-binding protein (TBP), and displays the characteristics of a molten globule. Limited proteolysis divides Myc1-167 in two halves, by cleaving in a predicted linker region between two hotspot mutation regions: Myc box I (MBI) and Myc box II (MBII). The N-terminal half (Myc1-88) is unfolded and does not alone bind to target proteins, whereas the C-terminal half (Myc92-167) has a partly helical fold and specifically binds both MM-1 and TBP. Although this might suggest a bipartite organization in the c-Myc TAD, none of the N and C-terminal fragments bind target protein with as high affinity as the entire Myc1-167, or display molten globule properties. Furthermore, merely linking the MBI with the C-terminal region, in Myc38-167, is not sufficient to achieve binding and folding properties as in Myc1-167. Thus, the entire N and C-terminal regions of c-Myc TAD act in concert to achieve high specificity and affinity to two structurally and functionally orthogonal target proteins, TBP and MM-1, possibly through a mechanism involving molten globule formation. This hints towards understanding how binding of a range of targets can be accomplished to a single transactivation domain.

Autoantibodies against amino acid 200-239 (p200) in the predicted leucine zipper region of the Ro52 protein are associated with congenital heart block, a potentially fatal condition that may affect fetuses of women with Ro52 autoantibodies. To allow detailed studies of the antibodies associated with congenital heart block, B-cell derived combinatorial antibody libraries from patients were screened for Ro52 and p200 specific antibody clones. Two human monoclonal anti-p200 antibody fragments, S3A8 and M4H1, were isolated and analysed with regard to VHand VL gene utilization, somatic mutations and binding properties. Both identified clones recognized recombinant and native intact Ro52, and reacted only with p200 in a set of related Ro52 peptides. The specificity and affinity was confirmed by biosensor measurements. Structural analysis of overlapping peptides revealed increased helicity in the p200 peptide compared to non-recognized peptides, indicating epitope conformation as essential for antibody binding. Both monoclonals produced punctate nuclear and diffuse cytoplasmic staining in human and mouse cell lines. The identified antibodies, which react specifically with the leucine zipper structure of Ro52, will be valuable in further exploration of the mechanisms operating during development of Ro52 antibody-associated congenital heart block.

The commonly used HIV-1 protease assays rely on measurements of the effect of inhibitions on the hydrolysis rate of synthetic peptides. Recently an assay based on surface plasmon resonance (SPR) was introduced. We have taken advantage of the fact that the SPR signal is proportional to the mass of the analyte interacting with the immobilised molecule and developed two new improved efficient competition assay methods. Thus, high molecular weight binders were used as amplifiers of the surface plasmon resonance signal. Linkers were attached by a Heck reaction to the para-positions of the P1/P1' benzyloxy groups of a linear C2-symmetric C-terminal duplicated inhibitor to enable (a) biotin labelling or (b) direct immobilisation of the inhibitor to the biosensor surface matrix. The interaction properties of a series of 17 structurally diverse inhibitors was assessed and compared to previously reported data. The most sensitive assay was obtained by immobilising the enzyme and amplifying the signal with an antibody, giving a detection range between 0.1 nM and 10 microM. Immobilisation of the inhibitor resulted in a stable and durable surface but a narrower detection range (1-100 nM). The two competition assays are anticipated to be very suitable for fast screening of potential HIV inhibitors.