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HybriFree Technology

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HybriFree Technology

Our HybriFree Technology is based on B-cell cloning and used for efficient discovery of monoclonal antibodies from immunized chickens and rabbits, ready for their immediate production in mammalian cells. HybriFree antibody discovery is rapid, flexible and effective. It can also be used for cloning antibodies from recovered patients’ PBMCs or from existing hybridomas.

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Hybrifree publication in bmc

Antibody discovery with HybriFree Technology

ROBUST
DISCOVERY
PLATFORM

ROBUST TECHNOLOGY PLATFORM

Our platform proceeds from B-cells to CHO cells. Therefore, by not switching between different systems, we don't lose the antibodies.

UNIVERSAL

UNIVERSAL

Efficient antibody cloning from different species (such as human, rabbit, chicken, mouse, macaque, sheep, dog).

SEAMLESS
TRANSITION

SEAMLESS TRANSITION

From discovery to large scale production.

HIGH AFFINITY
ANTIBODIES

HIGH AFFINITY ANTIBODIES

Even without affinity maturation.

INTEGRAL
MEMBRANE
PROTEINS?

YES!

90%
SUCCESS RATE

90% SUCCESS RATE

You can't beat biology. HybriFree platform does it in 90% of the cases.

Successful antibody discovery requires good antigen. 

Icosagen has over 15 years of experience in recombinant 

protein production and antigen design that combined with 

HybriFree discovery platform, results in high quality antibodies.

Case study: Development of SARS-CoV-2 antibodies

Recruit patients recovered from Covid-19
Isolate SARS-CoV-2 neutralizing antibodies
Identify lead candidate(s)
Manufacturing (GMP) Clinical trials

Antibody discovery with HybriFree technology

Virus neutralization potency of patient serum samples

Total of 187 convalescent blood donoros:

  • Positive PCR test
  • Mild to moderate symptoms
  • Some hospitalized, but not in ICU

 

Criteria for antibody cloning:

  • Tested ACE2 blocking efficacy and live virus neutralization potency of patient serum

 

Patient with high viral neutralization potency were selected for antibody cloning

23G7 binds RBD in closed conformation and blocks ACE2 interaction

Modelling with two 23G7 Fabs bound to the tri-S protein indicated sterical interference of the Fabs limiting the unbound RBD to move in the "up" ACE2 binding conformation

23G7 is effective in therapeutic and prophylactic SARS-CoV-2 challenge in Syrian Golden Hamsters

Development of humanized SARS-CoV-2 antibodies

1. Immunization of rabbits with the SARS-CoV-2 S1 protein (Wuhan)
2. Development of antibodies via HybriFree platform
3. Screening against VoC (Variant of concern) to identify cross-neutralizing clones
4. Sequence humanization

79C3 clone was identified as potent SARS-CoV-2 VoC neutralizing antibody. Humanization on 79C3 retained developability and functional neutralization profile.

Summary of SARS-CoV-2 case study

We identified highly potent SARS-CoV-2 neutralizing antibodies that were active at low picomolar concentrations

  • We identified the structural characteristics of our lead 23G7 antibody molecule
  • 23G7 antibody elicited effective virus neutralizing efficacy in Syrian Golden Hamster models
  • We identified VoC neutralizing antibodies

 

Formatting IgG into secretory IgA results in increased neutralization activity and resistance to independent mutations

Nebulized antibody delivery through inhalation results in broad respiratory tract distribution that leads to significantly reduced viral load in NHP models

HIGHLY POTENT
SARS-CoV-2
NEUTRALIZING
ANTIBODIES

HIGHLY POTENT SARS-CoV-2 NEUTRALIZING ANTIBODIES

We identified highly potent SARS-CoV-2 neutralizing antibodies that were active at low picomolar concentrations

REDUCED VIRAL
LOAD IN NHP
MODELS

REDUCED VIRAL LOAD IN NHP MODELS

Nebulized antibody delivery through inhalation results in broad respiratory tract distribution that leads to significantly reduced viral load in NHP models

STRUCTURAL
CHARACTERISTICS
IDENTIFIED

STRUCTURAL CHARACTERISTICS

We identified the structural characteristics of our lead 23G7 antibody molecule

HybriFree in vivo B-cell cloning approach
generates antibodies with optimal biochemical properties
and built-in, excellent developability.

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