Method of producing bispecific molecules by protein trans-splicing
First Claim
1. A method of producing a bispecific molecule having a first antigen recognition portion that binds a C3b-like receptor and a second antigen recognition portion that binds a pathogenic antigenic molecule, comprising contacting an N-intein first antigen recognition portion and a C-intein second antigen recognition portion under conditions such that protein trans-splicing occurs, wherein said N-intein first antigen recognition portion comprises said first antigen recognition portion conjugated to the amino terminus of an N-intein of a split intein, and wherein said C-intein second antigen recognition portion comprises said second antigen recognition portion conjugated to the carboxy terminus of a C-intein containing a splice junction of said split intein, wherein the amino acid residue immediately at the C-terminal side of the splice junction of said C-intein is an amino acid residue selected from the group consisting of cysteine, serine, and threonine.
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Abstract
The invention provides methods of using protein trans-splicing for the production of bispecific molecule which has a first antigen recognition portion that binds a C3b-like receptor and a second antigen recognition portion that binds an antigenic molecule present in the circulatory system of a mammal. The invention also provides bispecific molecules produced by protein trans-splicing. The bispecific molecules of the invention can be used for the clearance of pathogenic antigenic molecules from the circulatory system of a mammal. The invention further provides methods of using protein trans-splicing for the production of polyclonal libraries of bispecific molecules, which comprise populations of bispecific molecules with different antigen recognition specificities. Such polyclonal libraries of bispecific molecules can be used for targeting multiple epitopes of a pathogenic antigenic molecule and/or multiple variants of a pathogenic antigenic molecule.
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Citations
30 Claims
- 1. A method of producing a bispecific molecule having a first antigen recognition portion that binds a C3b-like receptor and a second antigen recognition portion that binds a pathogenic antigenic molecule, comprising contacting an N-intein first antigen recognition portion and a C-intein second antigen recognition portion under conditions such that protein trans-splicing occurs, wherein said N-intein first antigen recognition portion comprises said first antigen recognition portion conjugated to the amino terminus of an N-intein of a split intein, and wherein said C-intein second antigen recognition portion comprises said second antigen recognition portion conjugated to the carboxy terminus of a C-intein containing a splice junction of said split intein, wherein the amino acid residue immediately at the C-terminal side of the splice junction of said C-intein is an amino acid residue selected from the group consisting of cysteine, serine, and threonine.
Specification