Engineered Rabbit Antibody Variable Domains and Uses Thereof

A method for producing a rabbit VL or VH domain that immunospecifically binds to an antigen, wherein said method is a first method comprising:

• a) selecting from a phage expression library a set of DNA sequences that encode VH or VL domains that immunospecifically bind to said antigen, said phage expression library prepared from DNA or cDNA sequences that encode VH or VL domains and that were obtained from a biological sample of a rabbit containing cells that express immunoglobulins;
• and
• b) expressing said set of sequences, or a subset thereof, in bacteria as a fusion protein with CAT, selecting bacteria that have chloramphenicol resistance by virtue of CAT expression, and obtaining sequences encoding rabbit VH or VL domains from the selected bacteria;
• or a second method comprising:
• a) expressing sequences of DNA or cDNA that encode VH or VL domains in bacteria as a fusion protein with CAT, wherein said DNA or cDNA sequences were obtained from a biological sample of a rabbit containing cells that express immunoglobulins;
• b) selecting bacteria that have chloramphenicol resistance by virtue of CAT expression, and obtaining the set of sequences encoding rabbit VH or VL domains from the selected bacteria;
• and
• c) selecting from a phage expression library DNA sequences that encode VH or VL domains that immunospecifically bind to said antigen, wherein said library is prepared from said set of sequences, or subset thereof.