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Developability Assessment & Optimization

GenScript's provides chemical stability, physical stability

and best solution properties to the target clones

Peptides and proteins have significant potential in therapeutic niche, but failures still happen and are mostly related to intolerable toxicity, low efficacy and undesired biophysicochemical attributes. The attrition rate of the therapeutic drug could be further reduced by "developability" at an early stage. Developability is an analysis of the suitability of a biological lead molecule to be successfully developed into an effective therapy. Developability Assessment and optimization contributes to streamline the potency, safety and manufacturability. Developability screening allows developable candidates to be very effective, minimally toxic, and to reach the preclinical and clinical use.

GenScript`s Developability Assessment & Optimization platform provides chemical stability, physical stability and best solution properties to the target clones. We provide formulation mitigation strategies to avoid stability risks with solubility, hydrolysis, oxidation, aggregation, adsorption, denaturation and contaminations. Our services assess the targets using bioinformatics in silico methods, degradation and oxidation analysis by size exclusion chromatography (SEC) Biacore T200, PTMs, colloidal and thermal stability using high-throughput methods, such as CE-SDS, LC-MS analysis.

  • Manufacturability Assessment

    - PTMs
    - Chemical stability
    - Physical Stability

  • Safety
    Assessment

    - In Silico modeling
    - Immunogenicity

GenScript`s Developability Assessment and Optimization Conditions

Category Item Description Solution
Biophysical characterization and improvement Thermostability
  1. Incubate at 40 degrees up to 4 weeks, aliquots are analyzed by SEC-HPLC or ELISA.
  2. Incubate at gradient temp. up to  70 degrees 20 min,aliquots are analyzed by SEC-HPLC or ELISA.
  3. DSC
  1. Optimizing hydrophobic core and charge cluster residues
  2. Optimizing conserved residues
  3. Removing hydrophobic surface residues
Aggregation Analyzed by SEC-HPLC
Freeze and thaw stability Up to five  freeze and thaw cycles, aliquots are analyzed by SEC-HPLC
Biomatrix stability Analyzed by  ELISA (serum, plasma) Optimizing Ab through high throughput screening
Expression and Solubility improvement Analyzed by ELISA or SEC-HPLC Removing/ reducing  hydrophobic surface residues
Chemical Stability & PTMs Asparagine Deamidation
  1. Validated via peptide mapping  followed by LC-MS/MS,
  2. Binding analysis by SPR  or FACS
  3. SEC-HPLC
 Optimized Ab through antibody engineering  should retain the same affinity and activity as reference Ab
Aspartate isomerization
Tryptophan oxidation
Hydrolysis
N-glycosylation

GenScript`s Developability Assessment Tools

Developability assessment & Optimization tools are useful to reduce attrition rates that help us to produce most innovative biological drug candidates. GenScript`s developability service provides the analysis of biochemical and biophysical properties including base on the prediction and characterization tools. The in silico homology modeling bioinformatics tools provide you the most important information with manufacturability and safety of drug targets.

  • Peptide mapping and LC-MS analysis

  • Potential PTMs in antibody
     

  • Agilent HPLC/UHPLC 1200/1260/1290

Case Studies for Developability Assessment and Optimization

  1. Identification of antibody stability using SEC- HPLC and the stability of an antibody at various temperatures (Room temperature (RT), 30℃, 40℃, 50℃ and 60℃ Read More »

  2. MS analysis for humanized Abs under forced oxidation with AAPH-24h oxidation, LC-MS analysis , Coverage>95% Read More »

    3. Clones Chains Sequence Modifications Modification Percent XIC Area
    AH01695 control　 1 YNEMFTGRVTL 　 　 1.62E+06
    1 YNEMFTGRVTL Oxidation@4(63) 8.913 1.58E+05
    1 DTLMISR 　 　 1.35E+06
    AH01695 AAPH-24h 1 YNEMFTGRVTL 　 　 1.18E+06
    1 YNEMFTGRVTL Oxidation@4(63) 70.898 2.87E+06
    1 DTLMISR 　 　 5.67E+05
    1 DTLMISR Oxidation@4(250) 26.143 2.01E+05

  3. PTM analysis of AH01674 shows the carbamidomethylation and deamidation modifications and percentage of the modifications Read More »

    4. One identified PTM hotspot M63 existing in AH01674 did not impact the binding activity and homogeneity under forced oxidation status. No predicted deamidation hotspot existed in both AH01674, and the experimental data confirmed the prediction. Optimized lead was selected for further in vivo animal efficacy study and commercial production stable cell line generation.

  4. In vivo animal study shows the wild-type IgG and the chimeric IgG – AS04498 shows no toxicological effects and the affinity is about 38 fold increase compared to wild-type IgG Read More »

    5. Ligand ka (1/Ms) kd (1/s) KD (M) Rmax (RU) Chi² (RU²) Fold increase
    Wild-type IgG 6.35E+04 0.005986 9.43E-08 121.3 0.263 1
    AS04498 5.77E+04 1.44E-04 2.50E-09 129.4 0.336 38