Glycosylated biopharmaceuticals are important in the global pharmaceutical market. Despite the importance of their glycan structures, our limited knowledge of the glycosylation machinery still hinders controllability of this critical quality attribute. To facilitate discovery of glycosyltransferase specificity and predict glycoengineering efforts, here we extend an approach to model biosynthetic pathways for all measured glycans, and the Markov chain modeling is used to learn glycosyltransferase isoform activities and predict glycosylation following glycosyltransferase knock-in/knockout. We apply our methodology to four different glycoengineered therapeutics (i.e., Rituximab, erythropoietin, Enbrel, and alpha-1 antitrypsin) produced in CHO cells, along with o-glycosylation and lipid profiles. Our models accurately predict N-linked glycosylation following glycoengineering and further quantified the impact of glycosyltransferase mutations on reactions catalyzed by other glycosyltransferases. By applying these learned GT-GT interaction rules identified from single glycosyltransferase mutants, our model further predicts the outcome of multi-gene glycosyltransferase mutations on the diverse biotherapeutics. We further apply this to study differential O-glycosylation and lipidomics. Thus, this modeling approach enables rational glycoengineering and the elucidation of relationships between glycosyltransferases and other enzyme classes, thereby facilitating biopharmaceutical research and aiding the broader study of glycosylation to elucidate the genetic basis of complex changes in glycosylation and the lipidome.

The present invention relates to a method of obtaining an enriched population of a target polynucleotide using a synthetic single guide RNA (sgRNA) for an sgRNA-guided nucleic acid-binding protein, as well as to a method of obtaining a pool of target-irrelevant synthetic single guide RNAs (sgRNAs) for a sgRNA-guided nucleic acid-binding protein. Also provided is a target polynucleotide and sgRNAs obtainable by the methods of the invention. Further envisaged is a kit comprising a pool of sgRNAs obtainable by the method of the invention, and the use of a pool of sgRNAs obtainable by the methods of the invention.

Provided is a method for screening antibodies against a specific antigen epitope, including: Incubating antigens with incorporation of photocrosslinking amino acids(designated as mutant antigen) with antibody library under light irradiation with suitable wavelength and energy, and selecting antibodies that covalently crosslink with the mutant antigen; then the antibodies selected are subjected to affinity maturation against wild-type antigen, and then epitope-directed antibodies obtained.

Provided in this disclosure, in some embodiments, are methods and compositions for treating maple syrup urine disease (MSUD) and other conditions characterized by excessive branched-chain amino acids.

A method and apparatus are provided for designing sequence-defined biomolecules, such as proteins using a data-driven, evolution-based process. To design proteins, an iterative method founded on a combination of an unsupervised sequence-based model with a supervised functionality-based model can select candidate amino acid sequences that are likely to have a desired functionality. Feedback from measuring the candidate proteins using a high-throughput gene-synthesis and a protein screening process is used to refine and improve the models guiding the candidate selection to the most promising regions of the very large amino acid sequence search space.

The present invention relates to a method for producing a panel of cells (i.e. a cell library) expressing various different mutant variants of a protein of interest, wherein only one of said mutant variants is expressed per cell from a single gene copy. The present invention also relates to a method or cell library for identifying a mutant variant of a protein of interest having a different or modified biological activity as compared to the corresponding wild-type protein of interest. According to the present invention the identified mutant variant of a protein of interest may be applied for white biotechnology.

The present invention relates to a method for producing a panel of cells (i.e. a cell library) expressing various different mutant variants of a protein of interest, wherein only one of said mutant variants is expressed per cell from a single gene copy. The present invention also relates to a method or cell library for identifying a mutant variant of a protein of interest having a different or modified biological activity as compared to the corresponding wild-type protein of interest. According to the present invention the identified mutant variant of a protein of interest may be applied for white biotechnology.

The present invention relates to variant AAV capsid polypeptides, wherein the variant AAV capsid polypeptides exhibit increased transduction and/or tropism in human pancreatic tissue or human islets as compared non-variant parent capsid polypeptides.

Provided herein are methods, systems, and compositions for determining a base in a polynucleotide. In various aspects, the methods, systems, and compositions presented herein are useful for performing 4-base, 5-base, or 6-base sequencing of polynucleotide molecules, for example, from liquid biopsy samples or wherein the base is a low frequency mutation.

The present disclosure provides rapid and non-invasive methods for determining whether a patient exhibiting cancer symptoms, or at risk for hereditary cancers such as breast cancer, ovarian cancer, colon cancer, or skin cancer, will benefit from treatment with one or more therapeutic agents. These methods are based on detecting hereditary cancer-related mutations in small-volume dried biological fluid samples that are collected using a volumetric absorptive microsampling device (e.g., MITRA Tip). Kits for use in practicing the methods are also provided.