Archived Content

Cambridge Healthtech Institute’s Second Annual

Rapid Methods to Assess Quality & Stability of Biologics
Improving Prediction and Screening
Part of CHI’s 6th Annual The Bioprocessing Summit

August 18-19, 2014 | Renaissance Waterfront Hotel | Boston, Massachusetts

Day 1 | Day 2 | Short Courses | Download Brochure | Speaker Bios 

Tuesday, August 19

7:30 am Registration and Morning Coffee


7:55 Chairperson’s Remarks

NandaSubbaraoNanda Subbarao, Ph.D., Senior Consultant, Analytical CMC, Biologics Consulting Group




8:00 Methods for Rapid Assessment of Aggregation and Particle Formation

DeanRippleDean Ripple, Ph.D., Leader, Bioprocess Measurements Group, National Institute of Standards and Technology

Rapid assessment of the formation of particles in drug candidates requires the use of analytical methods that are suited to high throughput. Four main types of methods are considered: static and dynamic light scattering, static and dynamic microscopic imaging, temperature scanning methods with either calorimetric or fluorescent detection, and prospects for new, novel methods. For each method, I discuss the applicable size range, the sensitivity, and various advantages and disadvantages.

8:30 Emerging Methods for Measuring Sub Visible Particles

NandaSubbaraoNanda Subbarao, Ph.D., Senior Consultant, Analytical CMC, Biologics Consulting Group

The tools available for analysis of sub-visible particles in well-characterized protein products have increased over the past years in response to gradually increasing regulatory expectations to test for them. These emerging methods are based on different technologies. Therefore use of these methods together will provide a more complete description of the sub-visible particles, however the results cannot always be compared directly because they evaluate different features of the particles. The advantages and disadvantages of the different methods will be discussed.

9:00 Evaluation of the Stability of Low Concentration Maytansinoid ADCs in Infusion Bags and their Compatibilities with Administration Sets

Joyce Lin, Principal Analytical Associate, Analytical and Pharmaceutical Sciences, ImmunoGen, Inc.

Maytansinoid ADCs (AMCs) are used for the treatment of cancer. The AMC drug products are placed into infusion bags that contain appropriate diluent and administered intravenously. The stability of the AMCs and their compatibilities with the administration sets need to be evaluated before the start of the clinical trials. The human starting dose levels are relatively low and pose challenges during the assessment of compatibility of the diluted AMCs with infusion sets.

9:30 Q&A with Speakers

9:45 Coffee Break in the Exhibit Hall with Poster Viewing

10:30 Recent Advances in Monitoring Protein Aggregation Kinetics and Mechanisms with Simultaneous Multiple Sample Light Scattering (SMSLS)

WayneReedWayne F. Reed, Ph.D., Murchison Mallory Chair Professor of Physics, Department of Physics, Tulane University

SMSLS measured real-time aggregation kinetics of several proteins under thermal and stir stressors up to concentrations >0.100g/cm3. Arrhenius behavior is found for thermal data, but there is no relationship between aggregation rates, which vary by >106, and Tm and unfolding activation energy. Rates under stir are surprisingly similar. Stirring effects of enhanced air/liquid interface exposure vs. mechanical shear were separated. Thermal and stirring aggregation mechanisms are different. The appearance of particulates during aggregation was monitored.

11:00 Poly-Specificity as an Early Metric for Antibody Developability

EricKraulandEric Krauland, Ph.D., Senior Director, Antibody Discovery and Optimization, Adimab LLC.

The developability of antibody therapeutics is a historically overlooked aspect in the early discovery process. To this end, we demonstrate that a simple flow cytometry-based poly-specificity assay predicts poor CMC properties by correlation to validated characterization techniques. But unlike these assays, the flow cytometry poly-specificity assay is also compatible with active selection from large and diverse antibody mixtures. Selecting for CMC properties and target biology in the earliest discovery stages aims to improve the efficiency of the overall development process.

11:30 Selected Poster Presentation: Endotoxin Contamination in BioPharmaceuticals: Overcoming False Negative Results Induced by Endotoxin Masking

Johannes Reich, MSc, Institute of Physical and Theoretical Chemistry, University of Regensburg, Germany

Due to the fulminant physiological response endotoxin testing is mandatory in pharmaceutical production and product release of parenteral drugs. In order to solubilize certain active pharmaceutical ingredients, formulations contain surfactants (e.g. polysorbate) and buffer components (e.g. citrate), which can interact with endotoxin contaminations and prevent accurate detection (masking). We demonstrate that endotoxin masking depends on various parameters and provide dedicated approaches for a reliable detection of endotoxin contaminations.

12:00 pm Enjoy Lunch on Your Own

1:15 Session Break 


1:55 Chairperson’s Remarks


Wayne F. Reed, Ph.D., Murchison Mallory Chair Professor of Physics, Department of Physics, Tulane University




2:00 Alternative Methods for Quantifying Temperature- and Formulation-Dependent Aggregation Rates

ChristopherRobertsChristopher J. Roberts, Associate Professor, Department of Chemical & Biomolecular Engineering, University of Delaware

Reliably predicting protein aggregation rates from accelerated storage conditions remains an outstanding challenge for formulation scientists. Issues that need to be overcome include: sufficiently accurate means to quantify how rates change with storage condition and non-linear effects that make extrapolations difficult to perform accurately. This talk presents illustrative methods to improve predictions of aggregation rates, with monoclonal antibodies as case studies, and also highlights remaining challenges for future efforts.

2:30 The Measurement of KD at Low Concentration and Its Application as a High-Throughput Screening Technique for Protein-Protein Interaction Measurements

AnthonyYoungAnthony L. Young, Ph.D., Principal Scientist, Pharmaceutical Research and Development, Pfizer, Inc.

The light scattering measurement is routinely run in a high-throughput format to quickly determine the necessary diffusion coefficient versus concentration curves. The use of a robotic liquid handler can reduce the preparation time of the dilution sequence. This talk will cover the use of the liquid handler in combination with the dynamic light scattering instrument to generate KD values that are used to screen proteins and protein formulations for development. The data from several different protein isoforms will be discussed to illustrate the screening process and show typical data.

3:00 Simultaneous Stability and Aggregation Assessment by Isothermal Chemical Denaturation

Freire_ErnestoErnesto Freire, Ph.D., Professor, Biology and Biophysics, Johns Hopkins University

Stability and aggregation are two of the most important hurdles in the formulation of biologicals. Isothermal chemical denaturation (ICD) provides the most accurate way of measuring protein stability at room, physiological or storage temperatures under different solvent or formulation conditions, yielding reliable thermodynamic stability parameters. Furthermore, ICD experiments performed at different protein concentrations provide a quantitative assessment of protein aggregation in the native and denatured states. ICD is ideally suited to optimize the formulation of proteins hard to formulate, highly concentrated formulations, bispecific antibodies and antibody drug conjugates. In this presentation, the fundamentals of ICD and its application to the evaluation of protein stability and optimization of formulation conditions will be discussed.

3:30 Refreshment Break in the Exhibit Hall with Poster Viewing

4:15 Application of DSF as a High-Throughput Tool in Protein Characterization and Formulation Development

Shuai "Sunny" Shi, Ph.D., Senior Scientist, Sterile Product Development, Merck

In this study, we benchmarked DSF against the conventional thermal technique, differential scanning calorimetry (DSC), and more importantly made an attempt to predict protein thermal aggregation kinetics by DSF. We have defined three levels of correlations between DSF/DSC transition temperature and real-time thermal aggregation kinetics which will be shown in 3 individual case studies. We will also demonstrate the unique application of DSF in studying concentration-dependent thermal behaviors especially in the high-concentration range.

4:45 Increasing the Throughput of Protein Formulation Screening Using 96-Well Plate Format

QingyanHuQingyan Hu, Scientist, Ph.D., Scientist, Formulation Development, Regeneron, Inc.

To increase throughput during formulation screening, the use of a 96-well plate format was explored for candidate selection and formulation development. Multiple mAb candidates were screened against different buffer/pH and excipients using the 96-well plate format. In addition, the stability study results obtained using the 96-well plate format was compared to the results from using glass vials. With the incorporations of an automated liquid handling system and analytical instruments compatible with 96-well plates, this approach would greatly increase the throughput of formulation screening and development.

5:15 End of Conference

5:15- 6:00 Dinner Short Course Registration

6:00 – 8:30 Dinner Short Course*: Accelerated Stability Testing of Biologics 

*Separate registration required

Day 1 | Day 2 | Short Courses | Download Brochure | Speaker Bios