Bioassays for Biologics

Ready-to-Use, Cell-based Assays for Biologics Therapeutic Development Including Characterization, MOA Confirmation, nAbs Detection, Stability Studies, and Potency Testing

Bioassays are analytical methods used to measure a drug's biological activity or potency using a biological system representing the mechanism of action (MOA) of that drug. Eurofins DiscoverX® offers an extensive portfolio of cell-based, mechanistically-relevant bioassays, optimized and qualified with marketed biologic drugs or reference standards using ICH (International Council for Harmonization) guidelines. These robust, quantitative, ready-to-use cell-based assays use a simple, homogenous protocol. They can be easily implemented for characterization, potency measurement, neutralizing antibodies (nAb) detection, stability studies, and many other additional applications to interrogate biological functions and accelerate your biologics development program from discovery to post-market QC lot-release testing.

Bioassay

 

Read Blog All Targets List Biologics Target List

 

 

Advantages of Bioassays

Bioassays are ideal for determining drug potency and stability using a simple, homogeneous protocol that reflects the drug’s clinical MOA. These rapid cell-based assays bypass the need for expensive and time-consuming cell culturing or assay development, thereby potentially reducing timelines and errors.

 

  • Biologically Relevant – MOA-reflective, functional assays for monitoring and testing of biologic therapeutics
  • Qualified – Qualified with marketed biologic drugs or reference standards using International Council for Harmonization (ICH) guidelines
  • Robust & High Reproducibility – Superior intra-lot and inter-lot reproducibility and assay linearity for stability and potency testing and QC lot release applications
  • Simple Protocol, Fast Results – Easy-to-run, rapid homogeneous protocol amenable to implementation in multiple labs and high-throughput format for increased efficiency

 

Highlights of Bioassays
Benefits Outcomes Implementation
  • Decrease assay development and validation times
  • Provide accurate and precise data
  • Reduce costs
  • Accelerate potency testing and QC lot release
  • Obtain biologically relevant data with assays reflective of the target biology
  • Measure therapeutic potency
  • Interrogate appropriate MOA
  • Attain high lot-to-lot reproducibility
  • Simple, homogenous protocol that yields results in < 24 hours
  • Seamless method transfer for faster implementation at CMOs, CDMOs, or CROs
  • Amenable to miniaturization to run in high-throughput 384 well format

CMO = contract manufacturing company; CDMO = Contract Development and Manufacturing Organization; and CRO = Contract Research Organization

Eurofins DiscoverX Bioassays Save over 12 Months of Assay Development Time

Eurofins DiscoverX Bioassays Saves Over 12 Months of Assay Development Time

 

A quantitative and robust bioassay that is reflective of the drugs’ MOA is a critical component of any development program. Eurofins DiscoverX offers bioassays that can be implemented to accelerate your biologics development program from discovery to post-market. For characterization and QC lot-release testing, where assay reproducibility is paramount, ready-to-use (RTU) bioassays help you save over 12 months of assay development time.

 

Workflow

Workflow

 

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Kit Components

Kit Components

 

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Well-Structured Development Process Delivers Robust and Reproducible Assays

Well-Structured Development Process Delivers Robust and Reproducible Assays

A well-structured process for bioassay development yields assays that are robust and reproducible. The process begins with selecting the assay that most appropriately reflects the MOA of the drug, followed by optimization of assay parameters. Optimization through qualification steps includes refining conditions with the originator drug or reference molecule to establish robustness, precision, accuracy, and linearity of assay.

 

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Industry Requirements Drive Bioassay Development Strategy

Highlights of Bioassays
  • Reflective of MOA of the therapeutic or molecule
  • Stability-indicating
  • Relative potency range (50 to 150%)
  • Repeatable, robust, precise, and accurate
  • Reproducible
  • Intra-lot (plate-to-plate)
  • Inter-lot (lot-to-lot)
  • Analyst-to-analyst

Our dedicated team of Field Application Scientists ensures seamless and successful bioassay method transfer across all testing sites. Our analytical cell banking program is available to ensure long-term assay reproducibility and critical reagent supply assurance.

 

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MOA-Reflective Bioassays Deliver Highly Specific Responses

Bioassays measure highly target-specific response and enable interrogation of the therapeutic drug’s MOA.

Well-Structured Development Process Delivers Robust and Reproducible Assays

 

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Bioassays are ready-to-use cell-based assay kits with a simple, homogenous protocol and are provided as qualified or target-based. These bioassay kits deliver results in less than a day, enabling implementation in QC lot-release programs and transfer across multiple testing sites globally. Qualified bioassays have been optimized for their respective originator drug or reference standard, and can easily reduce assay development timelines by over 12 months. Ready-to-use target-based or MOA-based bioassays are optimized with a reference ligand. These assays can be readily implemented in development programs for biologic drug candidates that use the same MOA or specific target and can be further optimized and qualified with the clinical drug candidate.

 

Qualified Bioassays

Drug Name Qualified With Molecular MOA Assay Measures Qualification Data Bioassay Kits & Configuration
Rituximab - Cytotoxicity Antibody-Dependent Cellular-Cytotoxicity Request Data 97-1012Y026-00169 (2-plate)
97-1012Y026-00170 (10-plate)
Semaglutide Ozempic® GLP1R agonist cAMP Accumulation Request Data 95-0062Y2-00175 (2-plate)
95-0062Y2-00176 (10-plate)
Aflibercept Eylea® VEGF-Trap VEGFR2 (KDR) Dimerization View Data New HS Version*
93-0996Y1-00153 (2-plate)
93-0996Y1-00154 (10-plate)
Standard
93-0996Y1-00005 (2-plate)
93-0996Y1-00006 (10-plate)
Bevacizumab Avastin® Anti-VEGFA mAb VEGFR2 (KDR) Dimerization View Data New HS Version*
93-0996Y1-00165 (2-plate)
93-0996Y1-00166 (10-plate)
Standard
93-0996Y1-00001 (2-plate)
93-0996Y1-00002 (10-plate)
Ranibizumab Lucentis® Anti-VEGFA mAb VEGFR2 (KDR) Dimerization View Data New HS Version*
93-0996Y1-00167 (2-plate)
93-0996Y1-00168 (10-plate)
Standard
93-0996Y1-00003 (2-plate)
93-0996Y1-00004 (10-plate)
Adalimumab Humira® Anti-TNFα mAb IkB Degradation View Data 93-0538B15-00131 (2-plate)
93-0538B15-00132 (10-plate)
Anti-PD-1/PD-L1 Keytruda® PD-L1-induced PD-1 signaling SHP Recruitment View Data 93-1104Y19-00117 (2-plate)
93-1104Y19-00118 (10-plate)
Tocilizumab Actemra® Anti-ligand (IL-6) or anti-receptor (IL-6RA) drugs Ligand-induced receptor dimerization (IL6RA/IL6ST) View Data 93-1045B3-00109 (2-plate)
93-1045B3-00110 (10-plate)
Anakinra Kineret® IL-1RA (Inhibits IL-1) IL1R/IL1RAP Dimerization View Data 93-1032Y3-00105 (2-plate)
93-1032Y3-00106 (10-plate)
Darbepoetin Alfa Aranesp® EPOR agonist EpoR/EpoR Dimerization View Data 93-0965Y3-00019 (2-plate)
93-0965Y3-00020 (10-plate)
Epoetin Alfa Recombinant hEpo EPOR agonist EpoR/EpoR Dimerization View Data 93-0965Y3-00017 (2-plate)
93-0965Y3-00018 (10-plate)
Exenatide Byetta® GLP1R agonist cAMP Accumulation View Data 95-0062Y2-00101 (2-plate)
95-0062Y2-00102 (10-plate)
Liraglutide Victoza® GLP1R agonist cAMP Accumulation View Data 95-0062Y2-00099 (2-plate)
95-0062Y2-00100 (10-plate)
Insulin Glargine USP Insulin INSRb agonist SH2 Recruitment View Data 93-0466Y3-00011 (2-plate)
93-0466Y3-00012 (10-plate)
Insulin Lispro USP Insulin INSRb agonist SH2 Recruitment View Data 93-0466Y3-00009 (2-plate)
93-0466Y3-00010 (10-plate)
Insulin USP Insulin INSRb agonist SH2 Recruitment View Data 93-0466Y3-00007 (2-plate)
93-0466Y3-00008 (10-plate)
Panitumumab ‎Vectibix® Anti-EGFR mAb EGFR/ErbB2 Dimerization View Data 93-1051Y3-00093 (2-plate)
93-1051Y3-00094 (10-plate)
Pertuzumab Perjeta® Anti-HER2-dimerization mAb ErbB2/ErbB3 Dimerization View Data 93-1042Y3-00095 (2-plate)
93-1042Y3-00096 (10-plate)
Teriparatide ‎Forteo® PTHR agonist cAMP Accumulation View Data 95-0118Y2-00057 (2-plate)
95-0118Y2-00058 (10-plate)
Somatotropin, Somatropin Recombinant hGH GHR agonist SH2 Recruitment View Data 93-0756Y3-00023 (2-plate)
93-0756Y3-00024 (10-plate)
Follitropin alfa Gonal-F FSHR agonist cAMP Accumulation View Data 95-0119Y2-00103 (2-plate)
95-0119Y2-00104 (10-plate)
Sargramostim Leukine® Recombinant GM-CSF (CSFR2 agonist) CSF2RB/CSF2R Dimerization View Data 93-1078Y3-00111 (2-plate)
93-1078Y3-00112 (10-plate)

*HS Bioassay kits have been further optimized to deliver reproducible results with lower variability.

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Target-Based Bioassays

Target Molecular MOA Assay Measures Bioassay Kits & Configuration
CD19, CD20, CD38 Cytotoxicity Antibody- Dependent Cellular-Phagocytosis 97-1012Y026-00179 (2-plate)
97-1012Y026-00180 (10-plate)
97-1009Y025-00171 (2-plate)
97-1009Y025-00172 (10-plate)
CD19, CD20, CD38 Cytotoxicity Antibody- Dependent Cellular-Cytotoxicity 97-1012Y026-00169 (2-plate)
97-1012Y026-00170 (10-plate)
97-1009Y025-00177 (2-plate)
97-1009Y025-00178 (10-plate)
IL7R/IL2RG IL-7R/IL-2RG Agonists Ligand-induced receptor (IL7R/IL2RG) Dimerization 93-0997Y13-00081 (2-plate)
93-0997Y13-00082 (10-plate)
IL1RL1/IL1RAP IL-1RL1/IL1RAP Antagonists Ligand-induced receptor (IL1RL1/IL1RAP) Dimerization 93-1067Y3-00079 (2-plate)
93-1067Y3-00080 (10-plate)
TrkA-p75 Agonist drugs SH2 Recruitment 93-0529Y3-00147 (2-plate)
93-0529Y3-00148 (10-plate)
MCHR1 Agonist drugs β-Arrestin Recruitment 93-0940Y3-00149 (2-plate)
93-0940Y3-00150 (10-plate)
GHSR Agonist drugs β-Arrestin Recruitment 93-0242Y3-00141 (2-plate)
93-0242Y3-00142 (10-plate)
LHCGR Agonist drugs cAMP Accumulation 95-0106Y2-00137 (2-plate)
95-0106Y2-00138 (10-plate)
MRGPRX2 Agonist drugs β-Arrestin Recruitment 93-0309Y2-00143 (2-plate)
93-0309Y2-00144 (10-plate)
AGTR1 AGTR1 Agonists β-Arrestin Recruitment 93-0312Y2-00125 (2-plate)
93-0312Y2-00126 (10-plate)
AGTRL1 AGTRL1 Agonists β-Arrestin Recruitment 93-0250Y2-00121 (2-plate)
93-0250Y2-00122 (10-plate)
BDKRB2 BDKRB2 Agonists β-Arrestin Recruitment 93-0189Y2-00113 (2-plate)
93-0189Y2-00114 (10-plate)
CNR1 CNR1 Agonists β-Arrestin Recruitment 93-0959Y2-00119 (2-plate)
93-0959Y2-00120 (10-plate)
C5AR1 Anti-ligand (anti-C5a) β-Arrestin Recruitment 93-0557Y2-00059 (2-plate)
93-0557Y2-00060 (10-plate)
CCR2 Antagonist drugs β-Arrestin Recruitment 93-0192Y2-00061 (2-plate)
93-0192Y2-00062 (10-plate)
CXCR5 Antagonist drugs β-Arrestin Recruitment 93-0514Y2-00063 (2-plate)
93-0514Y2-00064 (10-plate)
Exendin-4 Agonist drugs β-Arrestin Recruitment 93-0300Y2-00029 (2-plate)
93-0300Y2-00030 (10-plate)
FGFR4-β-Klotho Agonist drugs SH2 Recruitment 93-1060Y3-00089 (2-plate)
93-1060Y3-00090 (10-plate)
GPBAR1 Agonist drugs cAMP Accumulation 95-0049Y2-00139 (2-plate)
95-0049Y2-00140 (10-plate)
GLP1R Agonist drugs β-Arrestin Recruitment 93-0300Y2-00027 (2-plate)
93-0300Y2-00028 (10-plate)
GLP2R Agonist drugs β-Arrestin Recruitment 93-0572Y2-00133 (2-plate)
93-0572Y2-00134 (10-plate)
IGF1R Agonist drugs SH2 Recruitment 93-0505Y1-00069 (2-plate)
93-0505Y1-00070 (10-plate)
IL2RB/IL2RG/IL2RA Modified agonists; anti-IL2 drugs Ligand-Induced Receptor (IL2RB/IL2RG/IL2RA) Hetero-oligomerization 93-1003Y3-00091 (2-plate)
93-1003Y3-00092 (10-plate)
IL6R/IL6ST Anti-IL-6 mAbs Ligand-induced receptor (IL6RA/IL6ST) dimerization 93-1045Y3-00043 (2-plate)
93-1045Y3-00044 (10-plate)
IL10RA/IL10RB Anti-receptor (IL-10RA) drugs Ligand-induced receptor dimerization (IL10RA/IL10RB) 93-0985Y3-00135 (2-plate)
93-0985Y3-00136 (10-plate)
IL17RA/IL17RC Anti-IL-17 mABs Ligand-induced receptor (IL17RC/IL17RC) Dimerization 93-0999Y3-00053 (2-plate)
93-0999Y3-00054 (10-plate)
IL31RA/OSMRb Anti-ligand (IL-31); anti-receptor drugs Ligand-induced receptor (IL31RA/OSMRb) dimerization 93-1002Y3-00083 (2-plate)
93-1002Y3-00084 (10-plate)
MC4R Antagonist drugs β-Arrestin Recruitment 93-0211Y3-00085 (2-plate)
93-0211Y3-00086 (10-plate)
NPY2R (β-Arrestin) Agonist, Antagonist drugs β-Arrestin Recruitment 93-0212Y2-00065 (2-plate)
93-0212Y2-00066 (10-plate)
NPY2R (cAMP) Agonist, Antagonist drugs cAMP Accumulation 95-0077Y2-00055 (2-plate)
95-0077Y2-00056 (10-plate)
PD-1 Anti-PD-1, PD-L1 drugs SHP Recruitment 93-1104Y19-00117 (2-plate)
93-1104Y19-00118 (10-plate)
PDGFRA Antagonist drugs SH2 Recruitment 93-0823Y3-00071 (2-plate)
93-0823Y3-00072 (10-plate)
PDGFRB Antagonist drugs SH2 Recruitment 93-0493Y3-00073 (2-plate)
93-0493Y3-00074 (10-plate)
Parathyroid Hormone(PTH) Receptor Agonist drugs β-Arrestin Recruitment 93-0315Y2-00047 (2-plate)
93-0315Y2-00048 (10-plate)
SIRPa (-CD47) Agonists, Antagonist drugs SH2 Recruitment 93-1135Y19-00129 (2-plate)
93-1135Y19-00130 (10-plate)
SSTR2 Agonist drugs β-Arrestin Recruitment 93-0181Y2-00067 (2-plate)
93-0181Y2-00068 (10-plate)
TrkB-p75 Antagonist drugs SH2 Recruitment 93-0530Y3-00075 (2-plate)
93-0530Y3-00076 (10-plate)
TrkC Antagonist drugs SH2 Recruitment 93-0464Y3-00077 (2-plate)
93-0464Y3-00078 (10-plate)
VIPR2 Agonist drugs β-Arrestin Recruitment 93-0317Y2-00087 (2-plate)
93-0317Y2-00088 (10-plate)
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Eurofins DiscoverX utilizes the robust and flexible Enzyme Fragment Complementation (EFC) technology to design bioassays for interrogating specific MOA that reflect the therapeutic target’s biology. Select examples are shown below of the bioassays’ design to interrogate different MOAs.

Potency Testing

During the late stages of drug development, potency testing is an integral part for the assessment of a drug’s specific ability to produce a given result. Bioassays are the primary tool used for measuring drug efficacy during potency testing, enabling researchers to determine how a particular drug dose will react in a given biological system. Bioassays are also critical bioanalytical tools that enable the determination drug’s MOA. Cell-based assays help establish the relative potency of a product by comparing the biological activity of a drug’s MOA with a reference preparation (from the USP, WHO, or in-house reference standard).

Potency Testing

Potency testing of Eurofins DiscoverX bioassays. The PathHunter® Jurkat PD-1 Bioassay Kit was used to test for accuracy, precision, and dilutional linearity. A. Anti-PD-1 samples were tested at varying potencies (ranging from 50% to 150%) relative to a reference sample of 100% potency. The experiments were performed in triplicates in three independent experiments by two analysts over three days. RP = relative potency. Results demonstrate that the assay was found to be extremely accurate (101.2%) and precise (3.1%). B. To demonstrate dilutional linearity of anti-PD-1 sample measurements, we analyzed “measured” against “expected” relative potencies and calculated the slope of the curve and the associated R2 values, arriving at an almost perfect R2 value of 99.4%.

Stability Testing

Stability Testing

Stability testing of Eurofins DiscoverX bioassays. A. Heat-stressed samples of an anti-PD-1 antibody were tested to evaluate if the PathHunter PD-1 Signaling Bioassay will differentiate the stressed material from the reference sample. A marked difference between the two response curves of the heat stress and reference samples was observed. B. To demonstrate that the PathHunter PD-1 Signaling Bioassay is stability-indicating, an additional set of experiments were performed as a part of a qualification study at BioOutsource with Nivolumab. The results indicate that only the heat-treated sample showed significant reduction activity, while the freeze thawed sample displayed no differences with respect to the untreated reference material. To further confirm these results, the heat-treated samples were run at 143% Relative Potency to assess the sensitivity to the assay when run at a different potency interval. At 143%, the results returned with a similar shift in the potency of the heat-treated sample as at 100%.

Neutralizing Anti-Drug Antibody Detection

Eurofins DiscoverX bioassays are widely used to detect neutralizing anti-drug antibodies (ADAs) or neutralizing antibodies (NAbs) in patient serum samples (e.g., up to 100% human serum), with high sensitivity and reproducibility.

Neutralizing Anti-Drug Antibody Detection

Example of detection of ADAs activity in the PathHunter Bevacizumab Bioassay Kit. Samples prepared in 10% pooled normal human serum. Increasing concentrations of NAbs against bevacizumab returns assay signal to levels comparable to VEGF alone. Further, the assay is sensitive enough to detect >100 ng of NAb when prepared in 10% human serum. FDA guidelines specify minimum sensitivity of 1000 ng/mL for detection of NAb in human serum.

Bioassay Programs

  • Analytical Cell Banks – Ensure long-term assay reproducibility with the production of bioassay cells that are considered as critical reagents from dedicated, well-characterized, and established cell banks. Learn More
  • Bioassay Certification – This program is designed to qualify CRO/CDMOs to run Eurofins DiscoverX bioassays so we can guide biopharma clients on which CRO/CDMOs they can use for their downstream therapeutic efforts. Learn More

Key Resources