Deep mining of antibody phage-display selections using Oxford Nanopore Technologies and Dual Unique Molecular Identifiers.

Antibody phage-display technology identifies antibody-antigen interactions through multiple panning rounds, but traditional screening gives no information on enrichment or diversity throughout the process. This results in the loss of valuable binders. Next Generation Sequencing can overcome this problem. We introduce a high accuracy long-read sequencing method based on the recent Oxford Nanopore Technologies (ONT) Q20 + chemistry in combination with dual unique molecular identifiers (UMIs) and an optimized bioinformatic analysis pipeline to monitor the selections. We identified binders from two single-domain antibody libraries selected against a model protein. Traditional colony-picking was compared with our ONT-UMI method. ONT-UMI enabled monitoring of diversity and enrichment before and after each selection round. By combining phage antibody selections with ONT-UMIs, deep mining of output selections is possible. The approach provides an alternative to traditional screening, enabling diversity quantification after each selection round and rare binder recovery, even when the dominating binder was > 99% abundant. Moreover, it can give insights on binding motifs for further affinity maturation and specificity optimizations. Our results demonstrate a platform for future data guided selection strategies.

Caries classification and management in the context of the CariesCare International (CCI™) consensus: a clinical case study.

The objective of this clinical case study is to illustrate the caries management four-step structured process, leading to personalised interventions specific for each individual patient's risks and needs, according to CariesCare International, derived from the International Caries Classification and Management System (ICCMS) for clinical practice. An 18-year-old female was diagnosed with higher caries risk at the individual level, and with several caries lesions at different severity stages, some likely active and others likely inactive. A care plan was co-created with the patient and delivered to obtain optimal health outcomes. Several issues pertinent to patient-centred care are discussed, including caries management at the individual and the tooth surface level, the preservation of tooth structure, patient's caries risk management, and prevention and control of caries lesions. The patient's perspective is taken into account and the health outcome focus of the system is highlighted.