ProCode

THE PROCODE ADVANTAGE

ProCode uses a proprietary transport and detection capability that self edits for and eliminates aggregation. No other technology can eliminate antibody aggregation, a major limitation in our competitor's diversity libraries and the major contributor to false positives. ProCode based diversity libraries are of such high quality that selection and affinity maturation results in a few dozen hits rather than the thousands typical of display and related technologies which cannot eliminate antibody aggregation. ProCode starts with human frameworks that are optimized to eliminate aggregation and ProCode is also used to eliminate aggregation in heavy and light chain constant and Fc domains.

Two ProCode scFv Selection Configurations:

The ProCode Antibody Platform:

ProCode is a stand-alone technology representing a unique approach to screening a synthetic antibody library. Selection for the antibody can either take place within an intact bacterial cell that expresses the library simultaneously with the antigen target or through library display on the bacterial periplasmic membrane with target added exogenously. With ProCode, each antibody scFv is physically linked to a leader sequence that is transported through the twin-arginine transferase (TAT) membrane complex. By linking the target molecule to an antibiotic gene like β-lactamase, only cells expressing an antibody that binds to the target molecule fused to β-lactamase can survive in the presence of ampicillin. Alternative selection schemes can be used in membrane display formats as a natural extension of leader association with the membrane.

Video of how ProCode works by Co-expression of library and target.

Click here to watch a video about how procode works

Selection of Antibodies from Synthetic Libraries

Vybion's Antibody libraries are created by combining a highly diverse collection of synthetically-constructed randomized CDR sequences that are further diversified by random lengths using a unique proprietary technique. Further diversity rounds can be also generated by a number of techniques such as error prone PCR either directed at CDR or flanking sequences.

PROCODE OVERVIEW

  • Rapid screening of libraries accomplished in days
  • Multiple configurations to accommodate different targets
  • Selection of domain specific antibodies by coexpressing a domain with the library to drive selection
  • Optimal solubility leads to stable molecules for more efficient and cost effective manufacturing, formulation and delivery

References:

  1. Mining mammalian genomes for folding competent proteins using Tat-dependent genetic selection in Escherichia coli. Lim HK, Mansell TJ, Linderman SW, Fisher AC, Dyson MR, Delisa MP. Protein Sci. 2009 Oct 14. [Epub ahead of print]PMID: 19830686
  2. Discovery of amyloid-beta aggregation inhibitors using an engineered assay for intracellular protein folding and solubility. Lee LL, Ha H, Chang YT, DeLisa MP. Protein Sci. 2009 Feb;18(2):277-86.PMID: 19177561
  3. Versatile selection technology for intracellular protein-protein interactions mediated by a unique bacterial hitchhiker transport mechanism. Waraho D, DeLisa MP. Proc Natl Acad Sci U S A. 2009 Mar 10;106(10):3692-7. Epub 2009 Feb 20.PMID: 19234130
  4. Efficient isolation of soluble intracellular single-chain antibodies using the twin-arginine translocation machinery. Fisher AC, DeLisa MP. J Mol Biol. 2009 Jan 9;385(1):299-311. Epub 2008 Nov 1.PMID: 18992254
  5. Following the path of a twin-arginine precursor along the TatABC translocase of Escherichia coli. Panahandeh S, Maurer C, Moser M, DeLisa MP, Müller M. J Biol Chem. 2008 Nov 28;283(48):33267-75. Epub 2008 Oct 3.PMID: 18836181
  6. Laboratory evolution of fast-folding green fluorescent protein using secretory pathway quality control. Fisher AC, DeLisa MP. PLoS One. 2008 Jun 11;3(6):e2351.PMID: 18545653
  7. Genetic selection for protein solubility enabled by the folding quality control feature of the twin-arginine translocation pathway. Fisher AC, Kim W, DeLisa MP. Protein Sci. 2006 Mar;15(3):449-58. Epub 2006 Feb 1.PMID: 16452624
  8. Twin-arginine translocation of active human tissue plasminogen activator in Escherichia coli. Kim JY, Fogarty EA, Lu FJ, Zhu H, Wheelock GD, Henderson LA, DeLisa MP. Appl Environ Microbiol. 2005 Dec;71(12):8451-9.PMID: 16332834
  9. A little help from my friends: quality control of presecretory proteins in bacteria. Fisher AC, DeLisa MP. J Bacteriol. 2004 Nov;186(22):7467-73. Review. PMID: 15516557