Expression and secretion system

Abstract

The invention provides an expression and secretion system, and methods of using the same, for the expression and secretion of one fusion protein in prokaryotic cells and a second fusion protein in eukaryotic cells. Also provided herein are nucleic acid molecules, vectors and host cells comprising such vectors and nucleic acid molecules.

1 - 72 . (canceled) 
     
     
         73 . An antibody produced using a process comprising culturing a host cell transformed with a vector comprising a nucleic acid molecule encoding a first polypeptide, a second polypeptide, a signal sequence, and a coat protein or adaptor protein, wherein:
 (a) the first polypeptide comprises a variable light chain (VL) domain comprising a VL-HVR1, a VL-HVR2, and a VL-HVR3;   (b) the second polypeptide comprises a variable heavy chain (VH) domain comprising a VH-HVR1, a VH-HVR2, and a VH-VHR3;   (c) the signal sequence is functional in both a prokaryotic cell and an eukaryotic cell and is encoded by a nucleic acid sequence that is 5′ to the nucleic acid sequence encoding the first polypeptide and 5′ to the nucleic acid sequence encoding the second polypeptide, wherein the signal sequence is encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 11 (consensus mBIP sequence, X ATG AAN TTN ACN GTN GTN GCN GCN GCN CTN CTN CTN CTN GGN GCN GTN CGN GCN, wherein N=A, T, C or G, and wherein X=ATG, X=ATG ACC, or Xis absent), SEQ ID NO: 16 (Opt1, ATG ATG AAA TTT ACC GTT GTT GCT GCT GCT CTG CTA CTT CTT GGA GCG GTC CGC GCA), SEQ ID NO: 17 (Opt2, ATG ATG AAA TTT ACT GTT GTT GCG GCT GCT CTT CTC CTT CTT GGA GCG GTC CGC GCA), or SEQ ID NO: 18 (Opt 3, ATG ATG AAA TTT ACT GTT GTC GCT GCT GCT CTT CTA CTT CTT GGA GCG GTC CGC GCA); and   (d) the first polypeptide and the second polypeptide form a full-length antibody, wherein the first and/or second polypeptide is fused to the coat protein or the adaptor protein.   
     
     
         74 . The antibody of  claim 73 , wherein the process further comprises recovering the antibody expressed by the host cell. 
     
     
         75 . The antibody of  claim 73 , wherein the antibody is recovered from the host cell culture medium. 
     
     
         76 . The antibody of  claim 73 , wherein the nucleic acid sequence encoding the signal sequence is operably linked to the 5′ end of the nucleic acid sequence encoding the first polypeptide and the 5′ end of the nucleic acid sequence encoding the second polypeptide. 
     
     
         77 . The antibody of  claim 73 , wherein the VL domain and the VH domain are each linked to a utility peptide. 
     
     
         78 . The antibody of  claim 77 , wherein the VH domain is linked to a CH1 domain and the VL domain is linked to a CL domain. 
     
     
         79 . The antibody of  claim 78 , wherein the utility peptide is selected from the group consisting of an Fc, a tag, a label, and a control protein. 
     
     
         80 . The antibody of  claim 79 , wherein the VL domain is linked to a control protein and the VH domain is linked to the Fc. 
     
     
         81 . The antibody of  claim 73 , wherein the nucleic acid encoding the first polypeptide or the second polypeptide is fused to a synthetic intron, wherein the synthetic intron comprises a nucleic acid encoding the coat protein or adaptor protein. 
     
     
         82 . The antibody of  claim 81 , wherein the synthetic intron is located between a nucleic acid encoding a VH domain and a nucleic acid encoding an Fc or a hinge. 
     
     
         83 . The antibody of  claim 82 , wherein the synthetic intron further comprises a nucleic acid encoding a naturally occurring intron of IgG1. 
     
     
         84 . The antibody of  claim 81 , wherein the nucleic acid encodes an adaptor protein, and wherein:
 (a) the adaptor protein is a leucine zipper; or   (b) the adaptor protein comprises the amino acid sequence of SEQ ID NO: 8, 9, 12, 13, 14, or 15.   
     
     
         85 . The antibody of  claim 81 , wherein the nucleic acid encodes a coat protein, and wherein the coat protein is selected from the group consisting of pI, pII, pIII, pIV, pV, pVI, pVII, pVIII, pIX, and pX of bacteriophage M13, f1, or fd. 
     
     
         86 . The antibody of  claim 85 , wherein a first Fab fusion protein is expressed in prokaryotic cells and a second Fab fusion protein is expressed in eukaryotic cells. 
     
     
         87 . The antibody of  claim 86 , wherein the first Fab fusion protein is a Fab-phage fusion protein. 
     
     
         88 . The antibody of  claim 87 , wherein the Fab-phage fusion protein comprises VH/CH1 fused to the pill. 
     
     
         89 . The antibody of  claim 86 , wherein the second fusion is a Fab-Fc or a Fab-hinge-Fc fusion protein. 
     
     
         90 . The antibody of  claim 89 , wherein the Fab-Fc or the Fab-hinge-Fc fusion protein comprises VH/CH1 fused to an Fc. 
     
     
         91 . The antibody of  claim 78 , wherein the CH1 domain comprises a portion of a natural splice donor sequence. 
     
     
         92 . The antibody of  claim 79 , wherein the Fc comprises a portion of a natural splice acceptor sequence.