Method for isolation of soluble polypeptides

Abstract

Polypeptides with desirable biophysical properties such as solubility, stability, high expression, monomericity, binding specificity or non-aggregation, including monomeric human V H s and V L s, are identified using a high throughput method for screening polypeptides, comprising the steps of obtaining a phage display library, allowing infection of a bacterial lawn by the library phage, and identifying phage which form larger than average plaques on the bacterial lawn. Sequences of monomeric human V H s and V L s are identified, which may be useful for immunotherapy or as diagnostic agents. Multimer complexes of human V H s and V L s are also identified. The V H s and V L s identified may be used to create further libraries for identifying additional polypeptides. Further, the V H s and V L s may be subjected to DNA shuffling to select for improved biophysical properties.

What is claimed is: 
     
         1 .- 23 . (canceled) 
     
     
         24 . A polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NO:8-54. 
     
     
         25 . (canceled) 
     
     
         26 . A nucleic acid sequence that encodes a polypeptide as claimed in  claim 24 . 
     
     
         27 - 49 . (canceled) 
     
     
         50 . A multimer comprising at least two V H  antibody fragments selected from SEQ ID NOs:8-22, or at least two V L  antibody fragments selected from SEQ ID NOs:23-54. 
     
     
         51 . (canceled) 
     
     
         52 . A multimer comprising at least one V H  antibody fragment selected from SEQ ID NOs:8-22, and at least one V L  antibody fragment selected from SEQ ID NOs:23-54. 
     
     
         53 .- 96 . (canceled) 
     
     
         97 . A pharmaceutical composition comprising the polypeptide sequence of  claim 24  and a pharmaceutically suitable agent. 
     
     
         98 .- 99 . (canceled)