Industries: Pharma / Biotech

Development and Validation of Bioanalytical Assays for Biologics: Quantitation (PK) and Immunogenicity Assays

Course Director: Robert Dodge, Ph.D.

Course Description - Course runs 9:00 to 5:00 both days

Part 1: Quantitation Assays

Quantitation assays are critical for the development of biologics and biopharmaceuticals as well as accurate detection of protein biomarkers. Immunoassays for quantitation of protein drugs in biological matrices (ex. plasma, serum, tissue) generally take much longer to develop and are subject to a wider range of interferences than traditional xenobiotic small molecule quantitation assays by LC-MS/MS. Part 1 of this interactive two day course focuses on these critical assays and will take you step-by-step through the development of quantitation assays for proteins including monoclonal antibodies.

The course begins with basic development, highlighting technical requirements, regulatory requirements and industry trends. We start by detailing how to select and maintain critical reagents, then move to assay formats. This includes understanding physical processes behind the four parameter model and how to interpret curve shape in terms of assay optimization. Then we move into qualification and validation of the assays to support pre-clinical and clinical studies. Part 1 of this course ends with preparation of the assay for use in a bioanalytical laboratory, including validation, establishing robustness, determining acceptable and practical system suitability criteria and suggestions for long term maintenance of the assay.

Part 2: Immunogenicity Assays

Part 2 of this course starts with an overview of immunology followed by the requirements of an immunogenicity program, including the establishment of a tiered testing approach for screening, confirmation assay analysis, titer assays and neutralizing antibody assays. The course then goes on to cover basic development of these assays, highlighting how to develop and label critical antibody reagents, different assay formats (ex. bridge, sandwich), including an analysis of strengths and weaknesses of each format. This is followed by a review of current scientific guidelines on how to establish cut-points (with a hands on calculation workshop with sample data at the end of the course). Optimization of screening assays to improve sensitivity and minimize outliers is also reviewed. The sensitivity of various assays to drug (assay drug tolerance) will be discussed and an overview of available technologies and sample pre-treatment approaches will be reviewed. Then we move into validation of the assays to support pre-clinical and clinical studies. The course will end with concepts of risk based testing approaches.

Who Should Attend

This two-day, two-part course is designed for those who perform, supervise, manage, audit, or oversee the validation of bioanalytical assays in a GLP laboratory. This includes, but is not limited to, professionals in Analytical Development, Quality Control, Quality Assurance, Validation, Regulatory Affairs, Pharmaceutical Development, Clinical Development, Medical Development, and Research and Development groups.

This course will also be of interest to Biopharmaceutical Development Companies, Biosimilar Drug Development Professionals, Regulatory Assessors, Consultants, and Clinical Laboratories.

First Day

Session 1: Immunochemistry Basics

  • Types of assays overview
  • Antibody chemistry, class, and sub-type
  • Preparation and purification of monoclonal and polyclonal antibodies

Session 2: Assay Formats

  • Basic sandwich ELISA format
  • Alternative formats (ex. direct, competition)
  • Detection techniques (ex. absorbance, luminescence)
  • Multiplexing
  • Instrument platforms
  • Critical reagent qualification

Session 3: Modeling and Analyzing Data

  • Equilibrium equations
  • Linear, four and five parameter curves
  • Anchor Points
  • Troubleshooting assay performance using curve model results
  • Determining precision, accuracy and total error following white paper guidance
  • Normal distributions relevant to assay acceptance criteria

Session 4: Assay Validation

  • Test method SOPs
  • FDA and draft EMA guidance for assay validation
  • Current required parameters for assay validation
  • Fit for purpose biomarker validations
  • The validation report
  • Incurred sample reanalysis

Session 5: Practical Development and Validation Workshop

  • Review and troubleshooting of simulated development data
  • Calculating accuracy and precision of an assay with simulated data

Second Day

Session 6: Immunology

  • Basic immunology
  • Vaccines and protein drugs
  • Immunogenicity predication
  • IgE versus IgM/IgG immune response

Session 7: Testing Considerations

  • Pre-clinical versus clinical testing
  • Platforms
  • Case study of IgE response

Session 8:Bioanalytical Testing

  • Screening assay formats
  • Confirmatory assay
  • Titer assay
  • Neutralizing antibody assay
    • Overview
    • Examples
    • Functional versus effector function assays for antibody drugs

Session 9:Assay Design

  • Practical considerations
  • Positive controls
  • Statistical treatment and assay validation
  • Common interferences
  • Detection platforms

Session 10: Regulatory Guidelines for Validation

  • EMA guidance
  • FDA draft guidance
  • EMA draft guidance for monoclonal antibodies

Session 11: Risk Based Approach to Testing

  • Likelihood and consequences considerations
  • Historical antidrug antibody responses
  • Scientific white papers

Session 12: Assay Validation and Cut-Point Determination Workshop

  • Scientific white papers
  • Calculation software and methodology
  • Example calculations

Learning Objectives

Upon completion of this course, attendees will have a clear understanding of regulatory agency expectations for bioanalytical development, and will have gained the background knowledge necessary to effectively plan bioanalytical assay development and validation programs for both quantitation assays and PK studies. Additionally, participants will learn to develop immunogenicity assays for detecting anti-drug antibodies for both marketed products and products in clinical development. Attendees will develop expertise in writing protocols, reports performing calculations, and acceptance limits for bioassay method validation.

Participants will have acquired insight into how to avoid common development and validation pitfalls and be able to quickly discriminate compliant from non-compliant validation activities. In addition, attendees will gain practical experience in applying what they have learned during hands on problem solving and calculations workshops.

Testimonials

"This course was very comprehensive, practical and clear, given by a Course Director who has been there.'" Denis G., Group Leader & Sr. Research Officer, BRI-NRC
"It's great course. Useful for studying the basic issues of PK and immunogenicity assay;" Ekaterina K., Senior Researcher of Laboratory/ Immunological Methods, BIOCAD