null

Antibody phage display services

Home / Antibody production /Phage display

Accelerate your antibody development program with ProteoGenix' phage display services and get your purified recombinant antibodies in less than 2 months! Our libraries are made of various monoclonal antibody formats including Fab, scFv or VHH and species! Thanks to our high diversity human libraries, humanization is not necessary for your therapeutic antibody development and you benefit from our strong guarantees!

Why choose ProteoGenix' antibody phage display services?

Guaranteed phage display services
Binders guaranteed

We guarantee AT LEAST 3 unique binders against your antigen!

IP free phage display antibody production
IP free

You get the full ownership of the antibody sequence generated!

Antibody phage display libraries
Very high diversity phage display libraries

Our premium libraries contain at least 109 different variants!

Phage display antibody formats
scFv, Fab or VHH formats available

The variety of our libraries allows us to propose a wide range of formats to fit a wide range of applications.

Antibody phage display immune library
Wide variety of species available

Besides our naïve libraries, you can also choose to develop immune libraries from a species of your choice without limitation.

Fast monoclonal antibody generation
Save time!

From antigen to antibody in less than 7 weeks!

Animal free monoclonal antibody production
No animal use

Naïve library screening does not require any animal use.

antibody library screening
Buy your library

Buy our naïve libraries or your own immune libraries!

Naïve or immune phage display libraries?

Naïve libraries Immune libraries
Species Human, rabbit, alpaca No limitation except humans
Formats Fab, scFv, VHH Fab, scFv, VHH
Number of binders ++ +++
Affinity of binders ++ +++
Potential immunogenicity issues No Yes
Animal use No Yes
Timelines 6-7 weeks 15-21 weeks + 6-7 weeks
Price + ++

 

Interested in our naïve library screening service or in constructing an immune library ? Please feel free to contact our dedicated account manager.

Our antibody phage display service process

Antibody phage display library construction
Antigen procurement or design and production
  • Peptide/small molecules: conjugation to carriers
  • Protein production including gene synthesis
  • Cell overexpressing target protein

Immune library

Immune library

Naive library

Antibody phage display library construction
Immune library construction
  • PBMC isolation
  • RNA extraction and cDNA synthesis
  • VH and VL PCR amplification
  • Library construction and QC
Antibody phage display screening
Library screening and biopanning
  • Screening of naive or immune library against antigen
  • 4-6 rounds of biopanning
Single phage screening
ELISA screening of single phage binders
  • ELISA screening and validation until identification of at least 3-10 different binders
Antibody sequencing
DNA extraction & antibody sequencing

Our antibody phage display service content

Step Content Timelines Deliverables
Library construction (only for immune libraries)
  • Animal immunization and titer tests
  • Cell isolation (spleen, bone marrow, PBMC)
  • RNA extraction
  • cDNA synthesis
  • VH and VL PCR amplification
  • cDNA assembly
  • Phagemid synthesis and cloning
  • Library QC
15-21 weeks
  • Progress report
Library screening and biopanning
  • Screening of the naïve or immune library
  • 4-6 rounds of biopanning
6-7 weeks
  • Progress report
ELISA screening
  • Screening and validation by ELISA until at least 3-10 different binders are identified
  • Progress report
DNA extraction and antibody sequencing
  • Phage DNA extraction and antibody sequencing
  • Detailed report
  • DNA sequence of the best binder
    Full ownership of the sequences
Options available:
  • Additional screening of single phage clones by ELISA against a specific protein/peptide/small molecule/cell
  • Additional screening of single phage clones by WB
  • Additional screening of single phage binders by flow cytometry on cells
  • Assessment of the affinity of single phage binders
  • Panning against cells overexpressing antigen of interest with library depletion against control cell

Case study: antibody generation against a small molecule by phage display

Project requirements

One of our customers requested a human antibody against a small molecule.
The small molecule was conjugated to different carriers to achieve optimal coating.
Biopanning was performed using our human naïve library of high diversity.
A total of 192 single phage binders were screened and validated by ELISA.
10 antibody sequences were required.

Biopanning results

Rounds Phage input (pfu) Phage output (pfu)
Panning with antigen coating Panning with no coating (negative control)
1 2.0×1012 8.9×106 3.4×106
4 1.0×1012 4.0×107 2.5×106

There is an obvious enrichment in anti-antigen binders even if it is difficult to remove the binders to carriers (known phenomenon).

Polyclonal phage ELISA

Phage quantity (pfu/well) Round 3 Round 4
Antigen Control Antigen Control
6×1012 3.035 2.584 3.630 3.451
2×1012 2.798 1.709 3.308 3.007
6.7X1011 1.508 1.069 3.028 2.128
2.2×1011 0.5666 0.5024 2.976 1.885
7.4×1010 0.1712 0.1932 2.581 1.704
2.5×1010 0.08020 0.09040 2.033 1.317
8.2×109 0.06090 0.09140 1.931 1.054
2.7×109 0.01620 0.01740 0.7235 0.7526
9×108 0.01290 0.01460 0.2341 0.2432

Although there are many unspecific binders (known phenomenon), an obvious enrichment in anti-antigen binders is observed (data in bold).

Monoclonal phage ELISA of the 11 unique antibodies identified (unique means unique sequence)

Clones Ag-carrier 1 Ag-carrier 2 Ag-carrier 3 No coating Carrier 1 Carrier 2 Carrier 3
A6 4.698 4.743 4.466 0.118 0.100 0.092 0.117
A9 4.769 4.766 4.420 0.100 0.092 0.078 0.074
B1 4.716 4.804 4.452 0.353 0.584 0.422 0.187
B2 4.494 4.482 4.874 0.081 0.122 0.109 0.100
D1 4.862 3.989 4.540 0.170 0.488 0.313 0.123
D8 4.862 4.090 4.486 0.057 0.106 0.102 0.059
F4 4.214 3.352 5.306 0.051 0.110 0.123 0.059
H2 4.797 4.537 4.588 0.084 0.213 0.188 0.112
H11 4.722 4.604 5.049 0.103 0.218 0.262 0.138
1A11 4.407 4.528 4.562 0.054 0.086 0.115 0.079
1G1 4.956 4.381 5.218 0.076 0.114 0.138 0.077

We generated 11 unique clones that specifically and strongly bind to the small molecule antigen.
Our customer decided to express these 11 clones as recombinant antibodies and to order the screening of 192 additional single phages, which resulted in the identification of 7 new unique sequences.

Learn more about our capabilities of antibody phage display services. Discover our complete report by clicking on the “GET A PDF REPORT” button.

What is phage display?

The phage display technology involves the determination of interaction partners of a protein (protein-protein, protein-DNA or protein-peptide interactions), which would be used as the “bait” for phages expressing peptides, proteins or antibodies coded by a huge DNA library (consisting of all coding sequences of a given organism). The function of that protein may therefore be determined.

Phage display cycle: phage display library, panning/biopanning and validation by ELISA

The phage display technique uses the filamentous phage, a virus that lives on E. coli. This method has proven to be a powerful technique for the interrogation of libraries containing billions of different proteins or peptides.
The application of phage display on large antibody libraries has been a real success with regard to the isolation of monoclonal antibodies which are highly specific against an antigen.
These new methods using libraries composed of billions of antibodies expressed on phages offer highly valuable alternatives that have proven to be as effective as traditional methods in the generation of billions of components and their quick selection to identify the most interesting candidates for your research.

As such, phage display is a useful and state-of-the-art tool widely used in therapeutic antibody discovery.

Advantages of antibody phage display

The major advantage of phage display lies in the simplified structure, stability and therefore reliability of phages, making it possible to be applied to various surfaces but also to in vivo applications.
The main characteristic of phage display technique is the physical coupling between genotype and phenotype (phage will « display » the protein on its outside while containing the gene for the protein inside). This requires the artificial introduction of encoding DNAs into phages, explaining the large diversity of phage-based antibody libraries. The engineered antibody fragments which are generated by phage display at ProteoGenix are single chain variable fragment format (scFv), Fab-based antibody format presentation and single-domain antibodies (VHH).

M13 bacteriophage in phage display

All ProteoGenix’ antibody phage display libraries are made of the M13 filamentous phage, a virus that infects the bacterium E. coli. This bacteriophage is the most commonly used for phage display and will be appropriate for you research.

M13 is a lysogenic filamentous phage, comprised of a circular and single-stranded DNA encased in a thin flexible tube made up of about 2700 copies of a single protein called pVIII, the major coat protein, and about 5 copies of pIII, the minor coat protein, present at the ends of the tube. Infection with M13 plasmids is not lethal for bacteria and they are used for many recombinant DNA processes.