Secretion of antibodies without signal peptides from bacteria
View Patent ↗The present invention is directed generally to compositions and methods for obtaining secretion of antibodies or antigen-binding antibody fragments from prokaryotes without the need for a signal peptide through making use of mutant host strains with altered secretory properties. In particular, the invention provides host cells and methods for obtaining secretion of antibodies or antigen-binding antibody fragments from bacteria without the need for a signal peptide and provides diverse libraries of antibody sequence resulting from such methods. The invention additionally provides diverse libraries.
1. A method for producing an antibody, comprising the steps of:
(a) culturing an E. coli host cell comprising a prlA4 (SEQ ID NO: 1) phenotype and at least one expression vector comprising a polynucleotide molecule encoding a heavy chain polypeptide of the antibody and a polynucleotide molecule encoding a light chain polypeptide of the antibody, under conditions that express the polynucleotide encoding the heavy chain polypeptide and the polynucleotide molecule encoding the light chain polypeptide of the antibody, wherein at least one of the polynucleotides encodes a polypeptide lacking a signal peptide,
(b) secreting the heavy chain polypeptide and the light chain polypeptide across a cytoplasmic membrane,
(c) forming the antibody from the heavy chain polypeptide and the light chain polypeptide, and
(d) isolating said antibody.
2. The method of claim 1 , wherein the antibody is selected from the group consisting of a human antibody, a mouse antibody, a rat antibody, a rabbit antibody, a camel antibody, a sheep antibody, a chimeric antibody, a humanized antibody, a fusion antibody, and an epitope-focused antibody.
3. The method of claim 1 , wherein the polynucleotides encoding the heavy chain polypeptide and the light chain polypeptide are present on different expression vectors.
4. The method of claim 1 , wherein the polynucleotides encoding the heavy chain polypeptide and the light chain polypeptide are both present on the same vector.
5. The method of claim 1 , wherein the polynucleotide encodes the heavy chain polypeptide lacking a signal peptide.
6. The method of claim 1 , wherein the polynucleotide encodes the light chain polypeptide lacking a signal peptide.
7. The method of claim 1 , wherein the polynucleotides encode the heavy and light chain polypeptides lacking signal peptides.