RESUMO
Monoclonal antibodies (mAbs) represent a promising modality for the prevention and treatment of viral infections. For infections that initiate from the respiratory tract, direct administration of specific neutralizing mAbs into lungs has advantages over systemic injection of the same mAbs. Herein, using AUG-3387, a human-derived mAb with high affinity to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its various variants, as a model mAb, we formulated the mAb into dry powders by thin-film freeze-drying, confirmed that the AUG-3387 mAb reconstituted from the dry powders retained their integrity, high affinity to the SARS-CoV-2 spike protein receptor binding domain (RBD), as well as ability to neutralize RBD-expressing pseudoviruses. Finally, we showed that one of the AUG-3387 mAb dry powders had desirable aerosol properties for pulmonary delivery into the lung. We concluded that thin-film freeze-drying represents a viable method to prepare inhalable powders of virus-neutralizing mAbs for pulmonary delivery into the lung.
Assuntos
Anticorpos Monoclonais , Liofilização , Pós , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Anticorpos Monoclonais/química , Anticorpos Monoclonais/administração & dosagem , Administração por Inalação , SARS-CoV-2/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Neutralizantes/administração & dosagem , Humanos , Inaladores de Pó Seco , Aerossóis , Pulmão/metabolismo , COVID-19/prevenção & controle , Tratamento Farmacológico da COVID-19RESUMO
The utility of blue fluorescent protein (BFP) has been limited by its low quantum yield and rapid photobleaching. A library targeting residues neighboring the chromophore yielded a variant with enhanced quantum yield (0.55 versus 0.34), reduced pH sensitivity and a 40-fold increase in photobleaching half-life. This BFP, named Azurite, is well expressed in bacterial and mammalian cells and extends the palette of fluorescent proteins that can be used for imaging.
Assuntos
Biblioteca Gênica , Proteínas Luminescentes/biossíntese , Proteínas Recombinantes/biossíntese , Citometria de Fluxo , Medições Luminescentes , Proteínas Luminescentes/genética , Fotodegradação , Proteínas Recombinantes/genéticaRESUMO
Degenerate codon libraries are frequently used in protein engineering and evolution studies but are often limited to targeting a small number of positions to adequately limit the search space. To mitigate this, codon degeneracy can be limited using heuristics or previous knowledge of the targeted positions. To automate design of libraries given a set of amino acid sequences, an algorithm (LibDesign) was developed that generates a set of possible degenerate codon libraries, their resulting size, and their score relative to a user-defined scoring function. A gene library of a specified size can then be constructed that is representative of the given amino acid distribution or that includes specific sequences or combinations thereof. LibDesign provides a new tool for automated design of high-quality protein libraries that more effectively harness existing sequence-structure information derived from multiple sequence alignment or computational protein design data.
Assuntos
Códon , Técnicas de Química Combinatória , Biblioteca Gênica , Engenharia de Proteínas , Algoritmos , Sequência de Aminoácidos , Dados de Sequência Molecular , Método de Monte Carlo , Análise de Sequência de Proteína , SoftwareRESUMO
La severidad en el aumento del problema de lesiones de los trabajadores en los Estados Unidos llama a la intervención de nuevos actores en el área de prevención de riesgos. Es así como nuestra profesión tomoa cada día más fuerza en este campo debido a que podemos aportar de forma importante a lo que está siendo realizado por otros profesionales. Las nuevas generaciones de kinesiólogos en Chile, como physical therapists en USA tenemos la gran responsabilidad de ser pioneros en este creciente campo ocupacional