Humanization

Our humanization pipeline combines state-of-the-art computational modeling, in silico developability assessment, and structure-guided engineering within a fully integrated one-stop platform. By seamlessly integrating antibody design, optimization, and experimental validation, we provide one of Canada’s most comprehensive and efficient solutions for therapeutic antibody development.

Humanization Overview

Our humanization workflow leverages CDR grafting and structure-guided engineering to transfer antigen-binding regions from animal-derived antibodies onto optimized human antibody frameworks, preserving binding affinity while reducing immunogenicity and improving therapeutic compatibility.

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Seamless Computationally Accelerated Therapeutics

In collaboration with 9Bio Therapeutics, our antibody humanization platform combines advanced computational design, structure-guided engineering, and developability assessment to optimize therapeutic candidates while preserving affinity and functionality.

By integrating in silico analysis with experimental validation, we streamline the transition from discovery antibodies to clinically relevant molecules with improved stability, reduced immunogenicity, and enhanced manufacturability.

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We offer multiple antibody engineering and optimization pipelines tailored to the specific needs of each therapeutic program, ranging from rapid sequence-based screening approaches to advanced structure-guided and molecular dynamics-driven computational workflows.

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EnceinteBIO

Available Pipelines
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Humanization

Hyper-humanization

High-resolution Hyper-Humanization

Germline alignment & framework selection

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All Germlines

All Germlines

In silico CDR grafting

✓

✓

✓

Generation of back-mutation variants
(affinity, developability and immunogenicity optimization)

✓

✓

✓

Sequence liability assessment
(deamidation, oxidation, etc.)

✓

✓

✓

Static structural modeling  of CDR boundary mutational tolerance

✓

✓

Dynamic structural modeling
(confirming steric alignment)

✓

Production efficiency evaluation
(transient)

✓

✓

✓

Affinity measurement
(crude extracts)

✓

✓

✓

Purification and manufacturability evaluation

Optional

Optional

Optional

Affinity measurement
(purified antibodies)

Optional

Optional

Optional

Full report

✓

✓

✓

Step by Step Humanization Process

Step 1 – CDR Grafting

Our molecular dynamics and structural simulation workflows enable detailed analysis of antibody stability, flexibility, and intermolecular interactions over time, allowing us to identify potential developability liabilities early in the design process. In addition, early in vitro measurement of developability and affinity characteristics allows selections of the best candidates early in the development process.
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Base Pipeline

Hyper-Humanization

High-Resolution Humanization
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Multiple Alignments to Find Best Germlines for Humanization and selection of VH/VL Pairing
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Alignments to all human germlines and best germline pick based on CDR geometry and orientation conservation

Step 2 – Micro-Scale Production

In the second step of the pipeline, small production of antibodies will allow hands-on measurment of antibody properties. To generate quality data, we use HPLC-SEC, iQue®3 (HTP-Flow Cytometer) and ELISA assays.

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Measurements include:
Production Yields
VH/VL pairing efficiency
Aggregation
Fragmentation

These properties are excellent predictors of antibody developability, manufacturability, solubility and stability.

Step 3 – Binding Strength and Specificity

Pipeline to ensure antibody binding and specificity will be developped according to the client’s needs and goals. Detailed report will be provided.

Binding strength and specificity measure by ELISA or flow cytometry.

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Step 4 – High-Resolution Kinetic Analysis

High-resolution kinetic curves can be generated for selected antibodies, ensuring complete characterization of antibody variants.

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Instruments Available

Acuity Arc HPLC (HPLC-SEC)

  • Assess protein purity and stability
  • Up to 96 proteins per day
HPLC SEC instrument

iQue®3 (HTP-Flow Cytometer)

  • Assess binding and blocking capacities on cells or beads
  • Up to 384 samples in 30 minutes
  • 3 lasers (VBR) & 15 detection channels
  • Low sample / time consumption
  • Different kits available
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Sony SH800S (FACS)

  • Assess binding and blocking capacities on cells or beads
  • Cell sorting into tubes or 384 well plates (SH800S)
  • 4 lasers (VYBR) & 8 detection channels (SH800S)
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Carterra LSA (HTP-SPR) & Octet Red 384 (BLI)

  • Obtain binding kinetics (KD,ka,kd)
  • Up to 1152 clones (SPR) & 384 (BLI) per run
  • Epitope binning (384×384 pairwise interactions)
  • Epitope mapping
  • Screening of purified or crude protein extracts
  • Low sample / time consumption (SPR)
  • Up to 1600 proteins per week
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