OPTIMISED HEAVY CHAIN AND LIGHT CHAIN SIGNAL PEPTIDES FOR THE PRODUCTION OF RECOMBINANT ANTIBODY THERAPEUTICS

摘要

Antibodies of interest comprising a heavy chain signal peptide and/or a light chain signal peptide or combination thereof and compositions are described. The method of determining the signal peptide including creating data set of antibodies signal peptides, clustering, selecting and creating recombinant antibodies for enhanced expression and secretion are described.

主权项

1. An antibody of interest comprising a heavy chain signal peptide, wherein the heavy chain signal peptide comprises a N-domain, a H-domain and a C-domain, wherein the heavy chain signal peptide is bound to the antibody via the C-terminal end of the C-domain; wherein the H-domain comprises an amino acid sequence of formula (I):(SEQ ID NO: 277)Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13, wherein
Xaa6 is any one of Trp, Phe or Ile;Xaa7 is any one of Val, Ile or Leu;Xaa8 is Phe or Leu;Xaa9 is Leu or Val;Xaa10 is Val or Leu;Xaa11 is Ala;Xaa12 is any one of Leu, Ile or Ala;Xaa13 is any one of Phe, Leu, Pro or Ala; andwherein the amino acid sequence of formula (I) has optionally one or two mutations;provided that an antibody showing the lowest secretion is excluded, wherein the lowest secretion is determined by a) fusing polynucleotides encoding the heavy chain of the antibody of interest to a polynucleotide encoding the heavy chain signal peptide as defined herein to obtain different combinations of heavy chain signal peptides and antibody of interest; b) creating at least one expression vector comprising the polynucleotides defined under a); and c) quantifying secretion of the antibody of interest encoded by the vector referred to under b) after transfection into an expression system to determine the antibody with the lowest secretion.