Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products

Tree nut allergies are considered an important health issue in developed countries. To comply with the regulations on food labeling, reliable allergen detection methods are required. In this work we isolated almond-specific recombinant antibody fragments (scFv) from a commercial phage display library bypassing the use of live animals, hence being consistent with the latest policies on animal welfare. To this end an iterative selection procedure employing the Tomlinson I phage display library and a crude almond protein extract was carried out. Two different almond-specific scFv (named PD1F6 and PD2C9) were isolated after two rounds of biopanning, and an indirect phage ELISA was implemented to detect the presence of almond protein in foodstuffs. The isolated scFvs demonstrated to be highly specific and allowed detection of 40 ng mL?1 and 100 ng mL?1 of raw and roasted almond protein, respectively. The practical detection limit of the assay in almond spiked food products was 0.1 mg g?1 (110e120 ppm). The developed indirect phage ELISA was validated by analysis of 92 commercial food products, showing good correlation with the results obtained by a previously developed real-time PCR method for the detection of almond in foodstuffs. The selected phage clones can be affinity maturated to improve their sensitivity and genetically engineered to be employed in different assay formats.

A constraint-based approach to quality assurance in service choreographies.

Knowledge about the quality characteristics (QoS) of service com- positions is crucial for determining their usability and economic value. Ser- vice quality is usually regulated using Service Level Agreements (SLA). While end-to-end SLAs are well suited for request-reply interactions, more complex, decentralized, multiparticipant compositions (service choreographies) typ- ically involve multiple message exchanges between stateful parties and the corresponding SLAs thus encompass several cooperating parties with interde- pendent QoS. The usual approaches to determining QoS ranges structurally (which are by construction easily composable) are not applicable in this sce- nario. Additionally, the intervening SLAs may depend on the exchanged data. We present an approach to data-aware QoS assurance in choreographies through the automatic derivation of composable QoS models from partici- pant descriptions. Such models are based on a message typing system with size constraints and are derived using abstract interpretation. The models ob- tained have multiple uses including run-time prediction, adaptive participant selection, or design-time compliance checking. We also present an experimen- tal evaluation and discuss the benefits of the proposed approach.