Universal libraries for immunoglobulins

A universal library for a prototype immunoglobulin of interest, comprising:

• twenty single predetermined amino acid libraries consisting of one single predetermined amino acid library for each of the twenty naturally occurring amino acids, wherein each single predetermined amino acid library comprises mutated immunoglobulins of interest wherein a single predetermined amino acid has been introduced into one or more positions in the mutated immunoglobulin by walk-through mutagenesis, and wherein each single predetermined amino acid library comprises a group of subset libraries, the library including subset libraries comprising:
• a) a subset library comprising prototype immunoglobulin of interest,
• b) subset libraries comprising mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in one of the six complementarity-determining regions of the immunoglobulin, with one subset library for each of the six complementarity-determining regions, thereby totaling 6 subset libraries;
• c) subset libraries comprising mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in two of the six complementarity-determining regions, with one subset library for each of the possible combinations of two of the six complementarity-determining regions, thereby totaling 15 subset libraries;
• d) subset libraries comprising mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in three of the six complementarity-determining regions, with one subset library for each of the possible combinations of three of the six complementarity-determining regions, thereby totaling 20 subset libraries;
• e) subset libraries comprising mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in four of the six complementarity-determining regions, with one subset library for each of the possible combinations of four of the six complementarity-determining regions, thereby totaling 15 subset libraries;
• f) subset libraries comprising mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in five of the six complementarity-determining regions, with one subset library for each of the possible combinations of five of the six complementarity-determining regions, thereby totaling 6 subset libraries;
• and
• g) one subset library comprising mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in all of the six complementarity-determining regions,
• wherein each subset library that contains mutated immunoglobulins, comprises mutated immunoglobulins in which the predetermined amino acid is present at least once at every position in the complementarity-determining region into which the predetermined amino acid has been introduced.

A library for a prototype immunoglobulin of interest, comprising nucleic acids encoding mutated immunoglobulins of interest wherein a single predetermined amino acid has been substituted in one or more positions in one or more complementarity-determining regions of the immunoglobulin of interest, the library including subset libraries comprising:

• a) a subset library comprising nucleic acids encoding prototype immunoglobulin of interest,
• b) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in one of the six complementarity-determining regions of the immunoglobulin, with one subset library for each of the six complementarity-determining regions, thereby totaling 6 subset libraries;
• c) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in two of the six complementarity-determining regions, with one subset library for each of the possible combinations of two of the six complementarity-determining regions, thereby totaling 15 subset libraries;
• d) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in three of the six complementarity-determining regions, with one subset library for each of the possible combinations of three of the six complementarity-determining regions, thereby totaling 20 subset libraries;
• e) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in four of the six complementarity-determining regions, with one subset library for each of the possible combinations of four of the six complementarity-determining regions, thereby totaling 15 subset libraries;
• f) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in five of the six complementarity-determining regions, with one subset library for each of the possible combinations of five of the six complementarity-determining regions, thereby totaling 6 subset libraries;
• and
• g) one subset library comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in all of the six complementarity-determining regions,
• wherein each subset library that contains nucleic acids encoding mutated immunoglobulins, comprises nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid is present at least once at every position in the complementarity-determining region into which the predetermined amino acid has been introduced.

A universal library for a prototype immunoglobulin of interest, comprising:

• twenty single predetermined amino acid libraries consisting of one single predetermined amino acid library for each of the twenty naturally occurring amino acids, wherein each single predetermined amino acid library comprises nucleic acids encoding mutated immunoglobulins of interest wherein a single predetermined amino acid has been introduced into one or more positions in the mutated immunoglobulin by walk-through mutagenesis, and wherein each single predetermined amino acid library comprises a group of subset libraries, the library including subset libraries comprising:
• a) a subset library comprising nucleic acids encoding prototype immunoglobulin of interest,
• b) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in one of the six complementarity-determining regions of the immunoglobulin, with one subset library for each of the six complementarity-determining regions, thereby totaling 6 subset libraries;
• c) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in two of the six complementarity-determining regions, with one subset library for each of the possible combinations of two of the six complementarity-determining regions, thereby totaling 15 subset libraries;
• d) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in three of the six complementarity-determining regions, with one subset library for each of the possible combinations of three of the six complementarity-determining regions, thereby totaling 20 subset libraries;
• e) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in four of the six complementarity-determining regions, with one subset library for each of the possible combinations of four of the six complementarity-determining regions, thereby totaling 15 subset libraries;
• f) subset libraries comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in five of the six complementarity-determining regions, with one subset library for each of the possible combinations of five of the six complementarity-determining regions, thereby totaling 6 subset libraries;
• and
• g) one subset library comprising nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid has been substituted in one or more positions in all of the six complementarity-determining regions,
• wherein each subset library that contains nucleic acids encoding mutated immunoglobulins, comprises nucleic acids encoding mutated immunoglobulins in which the predetermined amino acid is present at least once at every position in the complementarity-determining region into which the predetermined amino acid has been introduced.