According to a first aspect of the invention, a method for the generation of a cell line is provided, comprising the steps of (a) providing a plurality of mammalian B cells, wherein each of the plurality of B cells comprises a transgenic genomic DNA sequence encoding a marker protein inserted into an endogenous immunoglobulin locus comprised in said B cell, and wherein the transgenic genomic DNA sequence is amenable to cleavage by a site directed nuclease, particularly Cas9; (b) replacing the transgenic genomic DNA sequence encoding a marker protein with a second transgenic DNA sequence encoding a protein of interest; (c) sorting B cells based on the presence or absence of the marker protein; and (d) collecting B cells in which the marker protein is absent.

The present disclosure relates to a method of protein structure and amino acid residue interaction prediction based on saturation suppressor mutagenesis screening of a protein of interest. The method of the instant disclosure can be adapted for multi-protein complexes, and is useful where crystal structure of a protein of interest is not available.

A system for continuous mutagenesis to facilitate directed evolution, the system including DNA polymerases carrying the novel K54E point mutation, and other point mutations including I709N, A759R, D424A (herein called K54E_LF Pol I) and this methods of use to produce and detect lines where mutagenesis is continuous and does not exhibit the usual decline in mutagenesis with sequential cloning.

A system for continuous mutagenesis to facilitate directed evolution, the system including DNA polymerases carrying the novel K54E point mutation, and other point mutations including I709N, A759R, D424A (herein called K54E_LF Pol I) and this methods of use to produce and detect lines where mutagenesis is continuous and does not exhibit the usual decline in mutagenesis with sequential cloning.

The invention relates to methods for isolating traffic-enhancing mutants of drug delivery proteins. In one embodiment, the invention provides a carrier for delivering a therapeutic agent to an organelle, comprising a polypeptide encoded by a mutant penton base gene. In another embodiment, the invention provides a method of enhancing trafficking to a cell by administering a composition comprising a penton base (PB) protein with one or more mutations that enhance cellular entry.

The present invention relates functional ligands to target molecules, particularly to functional nucleic acids and modifications thereof, and to methods for simultaneously generating, for example, numerous different functional biomolecules, particularly to methods for generating numerous different functional nucleic acids against multiple target molecules simultaneously. The present invention further relates to functional ligands which bind with affinity to target molecules, such as molecules used proteins or peptides, such as LL37.

The present invention relates functional ligands to target molecules, particularly to functional nucleic acids and modifications thereof, and to methods for simultaneously generating, for example, numerous different functional biomolecules, particularly to methods for generating numerous different functional nucleic acids against multiple target molecules simultaneously. The present invention further relates to functional ligands which bind with affinity to target molecules, such as molecules used proteins or peptides, such as LL37.

The invention provides efficient methods for combining single-substitution libraries of nucleic acids that span and encode proteins of interest and for selecting resultant mutant proteins after expression which have improved properties or characteristics.

Methods and compositions for determining and/or monitoring the immune state of an individual are provided.

Methods and compositions for determining and/or monitoring the immune state of an individual are provided.