IMMUNI T

Standard antidrug antibody (ADA) assays:

• Screening assay: Development and validation of a sensitive, specific, precise, reproducible and robust assay.
  
• Confirmatory assay: Show specificity of the antibodies for confirmation and elimination of possible false positives.
  
• Neutralization assay: Use of a bioassay or ligand binding assay.
  
• Characterization assay: Determination of the antibody class and subclass, concentration/titer, specificity to other related proteins or tissues, etc.
  

Immunogenicity prediction before the first human dosage can comprise various characterization tools:

• In silico: T and B cell epitope mapping, sequence homology
  
• In vivo: Repeated dose studies in relevant animal models
  
• In vitro / ex vivo: T and B cell epitope binding, PBMC/CD4+ T cells proliferation with presentation of antigen epitopes
  

Critical immunogenicity: assay parameters:

• Reagents: Positive and negative controls, detection
  
• Matrix effects: Sample type, untreated vs treated
  
• Cut-off point: Statistical analysis
  
• Qualification/Validation: Sensitivity, specificity, precision, etc.

Oligonucleotide therapeutics

An Oligonucleotide drug candidate requires both a sensitive and reliable bioanalytical method; not only for use in plasma samples, but also in solid tissues, such as liver, kidney, skin, or brain. Sensitive assays are required for the lower therapeutic doses that are currently being administered as more effective drug regimens with higher potency and targeted delivery are developed. The bioanalytical method has to be selected taking into account any chemical modifications in the structure as well as the functionality of the therapeutic oligonucleotides. Numerous classes of therapeutic oligonucleotides compounds exist, including:

• siRNA;
  
• Antisense oligonucleotides;
  
• Aptamers;
  
• Immunomodulatory oligonucleotides (IMOs);
  
• miRNA-blocking oligonucleotides;
  
• RNA decoys;
  
• Ribozymes and DNA enzymes.
  

Hybridization assays are currently one of the most sensitive quantitative methods available for the determination of oligonucleotides achieving limits of quantitation down to the pg/mL level in a variety of different biological matrices including solid tissues. A number of different hybridization assay formats exist depending on the nature of the oligonucleotide drug candidate, however all of the assays work by exploiting the highly specific binding of the complementary oligonucleotide probe to the drug candidate, via Watson-Crick base-pairing.

Hybridization assay services

Our research and development group supports method transfer, development, and validation of hybridization assays for the determination of oligonuclotide therapeutics, as well as the subsequent analyses of study samples in support of regulated pre-clinical, and clinical studies.

Development and validating assays:

• Pre-validation assay demonstrating good accuracy and precision and selectivity, as well as in-process stability;
  
• Validation of the hybridization assay in compliance with the current regulatory requirements and GLP. The following parameters will be assessed:
  
  • Accuracy and Precision;
    
  • Selectivity;
    
  • Specificity;
    
  • Stability of the study sample (In-process, Freeze/Thaw, Long-term storage) Stability of Stock solutions;
    
  • Prozone;
    
  • Incurred Sample Reanalysis (ISR) performed during sample analysis.;
    
• Optimization of the extraction procedure, selection of probes, and establishing the assay conditions;
  
• Method transfer and comparison of data between laboratories;
  

Sample Analyses

Sample analysis is conducted in compliance with the current Regulatory Guidance's and White Papers applicable to the quantitative determination of drug therapeutics in biological fluids using ligand binding assays. The final bioanalytical report will be in a format suitable for regulatory submission.

Methods include standard toxicity studies (STS) and additional immunotoxicity studies conducted as appropriate. Whether additional immunotoxicity studies are appropriate should be determined by a weight of evidence review of various factor(s).

Immunotoxicology analytical data packages may comprise:

• Organ weight, cell density
  
• Circulating immunoglobulins
  
• Cytokines profiles
  
• Immune reaction protein levels
  
• Immunophenotyping
  

Immunotoxicology functional data packages may include:

• Immune response associated proteins by FACS
  
• Natural killer cells assays
  
• T cell response assays
  
• Drug-antigen proliferative response
  
• Complement assays
  
• All cell function assays
  

Additional studies are often composed but not limited to : T cell dependant antibody response, immunophenotyping, natural killer cell assay and various other cell functions, including cell mediated immunity.

We have expertise and experience in:

• Standard and biological drugs and cell based immuno-therapies;
  
• Medical device bio-compatibility;
  
• Food and environment bio-immunosafety and immunotoxicology services;
  
• Safety, toxicity and efficacy issues in immunology;
  
• Predictive immuno toxicology: tangible data early in development of lead therapy with immunotoxicity profiling adapted to your product;
  
• Environmental biohazard;
  
• Standard in vitro immunotoxicology assays using analytical assays (ELISA, RIA), flow cytometry for phenotyping and cell physiology analysis;
  
• Evaluation of veterinary and agricultural residues in food : genetically modified organisms;
  
• Conventional immune assays for immune monitoring and for immunotoxicity.
  

We offer a comprehensive range of in vivo,ex vivo and in vitro approaches:

• Immunomarkers, functional immunology, in vivo and in vitro analytical and functional assays;
  
• Immunohistochemistry and cell-based functional assays in support of:
  
  • Discovery lead selection;
    
  • Preclinical toxicology;
    
  • Clinical safety and efficacy studies.
    
• Designing of vaccine toxicology studies;
  
• Various in vivo models, such as host resistance models (bacterial, viral, fungal and protozoal) and hypersensitivity models (such as local lymph node assay : LLNA).
  

We offer a custom tailored approach to answer your specific immune system questions through:

• Defining the best approach for immunotoxicology testing of your test article.
  

We also perform:

Methods & Process Development & Validation;

• Integrative analysis of variations in lymphoid tissues, immune cells populations and a wide array of mediators of immune response;
  
• Immune capacity monitoring for clinical research subjects and compromised patients;
  
• Immune profiling for the healthy individual through basic and specific immunocompetence evaluation.