Abstract: The present disclosure relates to methods and materials for determining an isoelectric point for a protein including, for example, a binding molecule such as an antibody. The isoelectric points may be used in methods for the preparation of proteins. Such methods may comprise identifying amino acid residues that are exposed on the surface of the protein in a sequence of amino acid residues of the protein, assigning a pKa value to the surface exposed amino acid residues, and calculating the isoelectric point of the protein from the pKa values assigned to the surface exposed amino acid residues. The methods of the present disclosure may be used for selecting and utilizing a buffer for purification of a protein, preparing a protein formulation, purifying a protein and/or stabilizing a protein in solution.

Abstract: The present disclosure relates to materials and methods for enhancing the binding affinity of antibodies by introducing a number of targeted amino acid changes in antibody variable domains. The present disclosure provides novel methods of flowcharted amino acid change cycles, of identifying residues as members of unique proximity groups and of determining the degree of conspicuousness of amino acid residues in antibody variable regions. The disclosed methods are useful for the selection of variable region amino acid residues that are candidates for change to produce antibody variable domains with enhanced affinity for their binding partners.

Abstract: The present disclosure relates to methods and materials for enhancing the binding affinity of an antibody by means of generating a library or an array of targeted amino acid changes (e.g., mutations) at one or more positions in an antibody variable domain. The present disclosure relates to libraries or arrays and their uses for enhancing antibody affinity. The present disclosure relates to methods and materials for mutagenesis, including for the generation of novel or improved antibody variable domains and libraries or arrays of mutant antibody variable domains or nucleic acids encoding such mutant or modified variable domains.

Abstract: Methods are described for identifying the amino acid residues of an antibody variable domain which may be modified without diminishing the native affinity of the domain for antigen while reducing its immunogenicity with respect to a hereterologous species and for preparing so modified antibody variable domains which are useful for administration to heterologous species. Antibody variable regions prepared by the methods of the invention are also described.

Abstract: The present invention provides purified and isolated polynucleotides encoding Type I ribosome-inactivating proteins (RIPs) and analogs of the RIPs having a cysteine available for disulfide bonding to targeting molecules. Vectors comprising the polynucleotides and host cells transformed with the vectors are also provided. The RIPs and RIP analogs are particularly suited for use as components of cytotoxic therapeutic agents of the invention which include gene fusion products and immunoconjugates. Cytotoxic therapeutic agents or immunotoxins according to the present invention may be used to selectively eliminate any cell type to which the RIP component is targeted by the specific binding capacity of the second component of the agent, and are suited for treatment of diseases where the elimination of a particular cell type is a goal, such as autoimmune disease, cancer and graft-versus-host disease.