Improved Recombinant Adeno-Associated Viral Vector Production via Molecular Evolution of the Viral Rep Protein.

In the dynamic field of gene therapy, recombinant adeno-associated viruses (rAAVs) have become leading viral vectors due to their safety, long-term expression, and wide-ranging cell and tissue tropism. With numerous FDA approvals and commercial products underscoring their potential, there is a critical need for efficient production processes to achieve high vector titers and quality. A major challenge in rAAV production is the efficient packaging of the genome into the viral capsid, with empty or partially filled capsids often representing over 90% of the produced material.

Structural characterization of surface immobilized platinum hydrides by sensitivity-enhanced Pt solid state NMR spectroscopy and DFT calculations.

Supported single-site platinum hydride compounds are promising heterogeneous catalysts for organic transformations. Few methods exist to describe the structures of single-site Pt catalysts with atomic resolution because of their disordered structures and low Pt loadings. Here, we study the compounds formed when bis(tri--butylphosphino)platinum, Pt(P Bu), is supported on dehydroxylated SiO or SiO-AlO.

Procollagen-lysine 2-oxoglutarate 5-dioxygenases are responsible for 5R-hydroxylysine modification of therapeutic T-cell bispecific monoclonal antibodies produced by Chinese hamster ovary cells.

We present a detailed mass spectrometric analysis of three 2 + 1 T-cell bispecific monoclonal antibodies (TCB mAbs), where an unexpected +15.9950 Da mass shift in tryptic peptides was observed. This modification was attributed to the occurrence of 5R-hydroxylysine (Hyl) using a hybrid LC-MS/MS molecular characterization and CRISPR/Cas9 gene deletion approach.

Prediction of polyspecificity from antibody sequence data by machine learning.

Antibodies are generated with great diversity in nature resulting in a set of molecules, each optimized to bind a specific target. Taking advantage of their diversity and specificity, antibodies make up for a large part of recently developed biologic drugs. For therapeutic use antibodies need to fulfill several criteria to be safe and efficient.

The reduction behavior of sulfurized polyacrylonitrile (SPAN) in lithium-sulfur batteries using a carbonate electrolyte: a computational study.

Lithium-sulfur batteries (LSBs) have attracted attention due to their high theoretical energy density. This and various other advantages, such as the availability and non-toxicity of sulfur, raise interest in LSBs against the background of the energy revolution. However, a polysulfide shuttle mechanism can adversely affect the electrochemical performance of the cell.

How Solid Surfaces Control Stability and Interactions of Supported Cationic Cu(dppf) Complexes─A Solid-State NMR Study.

Organometallic complexes are frequently deposited on solid surfaces, but little is known about how the resulting complex-solid interactions alter their properties. Here, a series of complexes of the type Cu(dppf)(L) (dppf = 1,1'-bis(diphenylphosphino)ferrocene, L = mono- and bidentate ligands) were synthesized, physisorbed, ion-exchanged, or covalently immobilized on solid surfaces and investigated by P MAS NMR spectroscopy. Complexes adsorbed on silica interacted weakly and were stable, while adsorption on acidic γ-AlO resulted in slow complex decomposition.

Rabbits transgenic for human IgG genes recapitulating rabbit B-cell biology to generate human antibodies of high specificity and affinity.

Transgenic animals incorporating human antibody genes are extremely attractive for drug development because they obviate subsequent antibody humanization procedures required for therapeutic translation. Transgenic platforms have previously been established using mice, but also more recently rats, chickens, and cows and are now in abundant use for drug development. However, rabbit-based antibody generation, with a strong track record for specificity and affinity, is able to include gene conversion mediated sequence diversification, thereby enhancing binder maturation and improving the variance/selection of output antibodies in a different way than in rodents.

Predicting healthcare utilization in youth with type 1 diabetes: The importance of family level variables.

Objective: Little is known about how family factors impacting treatment adherence in type 1 diabetes are directly associated with unplanned healthcare utilization (e.g., emergency room visits and hospital admissions).

CAR-J cells for antibody discovery and lead optimization of TCR-like immunoglobulins.

T-cell bispecific antibodies (TCBs) are a novel class of engineered immunoglobulins that unite monovalent binding to the T-cell receptor (TCR) CD3e chain and bivalent binding to tumor-associated antigens in order to recruit and activate T-cells for tumor cell killing. In vivo, T-cell activation is usually initiated via the interaction of the TCR with the peptide-HLA complex formed by the human leukocyte antigen (HLA) and peptides derived from intracellular proteins. TCR-like antibodies (TCRLs) that recognize pHLA-epitopes extend the target space of TCBs to peptides derived from intracellular proteins, such as those overexpressed during oncogenesis or created via mutations found in cancer.

Characterization and prediction of positional 4-hydroxyproline and sulfotyrosine, two post-translational modifications that can occur at substantial levels in CHO cells-expressed biotherapeutics.

Biotherapeutics may contain a multitude of different post-translational modifications (PTMs) that need to be assessed and possibly monitored and controlled to ensure reproducible product quality. During early development of biotherapeutics, unexpected PTMs might be prevented by identification and characterization together with further molecular engineering. Mass determinations of a human IgG1 (mAb1) and a bispecific IgG-ligand fusion protein (BsAbA) demonstrated the presence of unusual PTMs resulting in major +80 Da, and +16/+32 Da chain variants, respectively.

Sphinx: merging knowledge-based and ab initio approaches to improve protein loop prediction.

Motivation: Loops are often vital for protein function, however, their irregular structures make them difficult to model accurately. Current loop modelling algorithms can mostly be divided into two categories: knowledge-based, where databases of fragments are searched to find suitable conformations and ab initio, where conformations are generated computationally. Existing knowledge-based methods only use fragments that are the same length as the target, even though loops of slightly different lengths may adopt similar conformations.

Gender Differences in the Developmental Links Between Conduct Problems and Depression Across Early Adolescence.

Various developmental models have attempted to explain the relationship between antisocial behavior and depressive symptoms in youth, often proposing intermediary processes such as social and academic functioning. However, few studies have tested these developmental models fully, particularly in mixed gender samples. The current study strives to fill this gap in the literature, examining these processes in an early adolescent sample.

Length-independent structural similarities enrich the antibody CDR canonical class model.

Complementarity-determining regions (CDRs) are antibody loops that make up the antigen binding site. Here, we show that all CDR types have structurally similar loops of different lengths. Based on these findings, we created length-independent canonical classes for the non-H3 CDRs.

Efficacy of selective serotonin reuptake inhibitors and adverse events: meta-regression and mediation analysis of placebo-controlled trials.

Background: It has been suggested that the efficacy of antidepressants has been overestimated in clinical trials owing to unblinding of drug treatments by adverse events.

Aims: To investigate the association between adverse events and the efficacy of selective serotonin reuptake inhibitors (SSRIs).

Method: The literature was searched to identify randomised, double-blind, placebo-controlled trials of SSRIs in the treatment of major depression.

MoFvAb: Modeling the Fv region of antibodies.

Knowledge of the 3-dimensional structure of the antigen-binding region of antibodies enables numerous useful applications regarding the design and development of antibody-based drugs. We present a knowledge-based antibody structure prediction methodology that incorporates concepts that have arisen from an applied antibody engineering environment. The protocol exploits the rich and continuously growing supply of experimentally derived antibody structures available to predict CDR loop conformations and the packing of heavy and light chain quickly and without user intervention.

[Is there an Association between Prevention Campaign Knowledge and Sun Protection Behaviour of Parents for their Children? Results of a Parent Survey].

Aim: The aims of this study are to assess prevalence of awareness of sun protection campaigns among parents in Bavaria, Germany, to analyse the impact of sociodemographic factors on campaign knowledge and the association between parental campaign knowledge and sun protection behaviour in their children.

Methods: A cross-sectional survey was undertaken in 2010-2011 in Bavaria, Germany, with parents of 4,579 children aged 5-6 years (response rate 61%).

Results: Prevalence of knowledge of sun protection campaigns is 13% among parents in Germany and independent of sociodemographic factors.

The MOBI-Kids Study Protocol: Challenges in Assessing Childhood and Adolescent Exposure to Electromagnetic Fields from Wireless Telecommunication Technologies and Possible Association with Brain Tumor Risk.

The rapid increase in mobile phone use in young people has generated concern about possible health effects of exposure to radiofrequency (RF) and extremely low frequency (ELF) electromagnetic fields (EMF). MOBI-Kids, a multinational case-control study, investigates the potential effects of childhood and adolescent exposure to EMF from mobile communications technologies on brain tumor risk in 14 countries. The study, which aims to include approximately 1,000 brain tumor cases aged 10-24 years and two individually matched controls for each case, follows a common protocol and builds upon the methodological experience of the INTERPHONE study.

Structure-based prediction of asparagine and aspartate degradation sites in antibody variable regions.

Monoclonal antibodies (mAbs) and proteins containing antibody domains are the most prevalent class of biotherapeutics in diverse indication areas. Today, established techniques such as immunization or phage display allow for an efficient generation of new mAbs. Besides functional properties, the stability of future therapeutic mAbs is a key selection criterion which is essential for the development of a drug candidate into a marketed product.

A robust high throughput platform to generate functional recombinant monoclonal antibodies using rabbit B cells from peripheral blood.

We have developed a robust platform to generate and functionally characterize rabbit-derived antibodies using B cells from peripheral blood. The rapid high throughput procedure generates a diverse set of antibodies, yet requires only few animals to be immunized without the need to sacrifice them. The workflow includes (i) the identification and isolation of single B cells from rabbit blood expressing IgG antibodies, (ii) an elaborate short term B-cell cultivation to produce sufficient monoclonal antigen specific IgG for comprehensive phenotype screens, (iii) the isolation of VH and VL coding regions via PCR from B-cell clones producing antigen specific and functional antibodies followed by the sequence determination, and (iv) the recombinant expression and purification of IgG antibodies.

Determinants of inadequate parental sun protection behaviour in their children--results of a cross-sectional study in Germany.

Objective: Unprotected sun exposure especially during childhood is a risk factor for skin cancer. A combined use of sun protection measures is recommended to protect children. However, the prevalence and determinants for combined use have been scarcely studied in children.

Attention bias in youth: associations with youth and mother's depressive symptoms moderated by emotion regulation and affective dynamics during family interactions.

Recent research has highlighted the potential role of attention bias for emotional stimuli as a possible cognitive risk factor for depression in youth. However, differences in youth emotion regulation or maternal affect may moderate the association between maternal and youth depression and youth attention biases. The current study investigated the relationship between maternal and youth depressive symptoms and youth (aged 11-17 years) attention bias for sad and happy faces in 59 mother-youth dyads, examining whether positive and negative maternal affect observed during structured interaction tasks or youth emotion regulation tendencies moderated associations between maternal and youth depression and attention biases.

Examining the developmental interface of cortisol and depression symptoms in young adolescent girls.

Despite the substantial amount of data supporting a link between HPA-axis functioning and depression, the ontogeny of this association is not known. The aim of the present study was to contribute data on the developmental interface of HPA-axis functioning and depression in girls by testing associations between repeated measures of depression symptoms and cortisol levels in childhood and early adolescence. Girls (N=232) and their mothers, who were participating in a longitudinal study, were interviewed about depression symptoms annually from ages 9 to 12 years.

Family Check Up effects on adolescent arrest trajectories: Variation by developmental subtype.

This study examines the effect of the Family Check Up intervention on the probability of arrests from ages 12 to 17 years for youth following heterogeneous developmental trajectories of antisocial behavior. Latent Growth Mixture Modeling results supported the presence of three developmental trajectories of arrests, including a large group of youth with few police contacts, a smaller group of youth showing early onset and chronic arrests, and a group with adolescent-onset arrests. In line with hypotheses, effects of intervention were seen within the adolescent-onset group, but not in the early onset chronic arrest trajectory group, or those youth with little police contact.

Differential splicing generates new transmembrane receptor and extracellular matrix-related targets for antibody-based therapy of cancer.

Alternative splicing has been shown to be deregulated in cancer and a link to growth stimulation has been established. Here we describe transmembrane and extracellular matrix-related targets generated by alternative splicing with a restricted pattern of expression in normal tissues and a deregulated pattern of expression in cancer as possible targets for therapeutic intervention with antibody-related agents. We focus on isoforms of transmembrane and extracellular matrix proteins, such as CD44, Claudin 18, L1 cell adhesion molecule and epithelial cellular adhesion molecule, fibronectin, tenascin, osteopontin and versican as well as transmembrane tyrosine kinases, such as fibroblast growth factor receptors, epidermal growth factor receptor and receptor d'origin nantais.

Maternal depression and the heart of parenting: respiratory sinus arrhythmia and affective dynamics during parent-adolescent interactions.

Maternal depression is associated with problematic parenting and the development of emotional and behavior problems in children and adolescents. While emotional regulatory abilities are likely to influence emotional exchanges between parents and teens, surprisingly little is known about the role of emotion regulation during parent-child interactions, particularly in high-risk families. Respiratory sinus arrhythmia (RSA) has been widely linked to emotion regulatory abilities in recent research, and the current study investigated RSA and maternal depression in relation to dyadic flexibility, as well as mutuality of negative and positive affect displayed during three discussion tasks between 59 mother-adolescent pairs (age 11-17 years).