Bioassays for Biologics

Ready-to-Use Cell-Based Assays for Comparability and QC Lot Release Testing

Pharmaceutical companies are developing an increasing number of biologics, biosimilars and biobetters, thereby creating a need for MOA-reflective cell-based assays that can support multiple stages of the biopharmaceutical development pipeline. DiscoverX offers the largest menu of “Thaw & Use” Bioassay kits that have been qualified with marketed biologic drugs or reference standards. Bioassay kits are also available as target-specific cell-based bioassays that are easy to implement in any lab from development to QC/Lot release.
 
Bioassay
 

Accelerate Assay Development Time by 9 Months with Robust, Highly-Reproducible Bioassays

Accelerate Assay Development Time by 9 Months with Robust, Highly-Reproducible Bioassays
 

Advantages of Bioassay

Bioassays are ideal for determining drug potency and stability using a simple, mix-&-read protocol that reflects the drug’s clinical mechanism of action. These rapid cell-based assays bypass the need for expensive and time-consuming cell culture, reducing lab work errors. Additionally, bioassays decrease assay development and validation times, provide accurate and precise data, reduce costs, facilitate a simplified global method transfer, and ultimately accelerate potency testing and QC lot release.

 
  • Biologically-Relevant — MOA-reflective, functional assays for monitoring and testing of biologic therapeutics
  • High Reproducibility — Superior intra-lot and inter-lot reproducibility and assay linearity for stability testing
  • Robust Assays – Highly reproducible assays for potency and lot release applications for immunotherapy drug discovery and development
  • Simple Protocol, Fast Results — Easy-to-run, rapid homogeneous protocol amenable to implementation in multiple labs and high-throughput format for increased efficiency
 

Features of Ready-to-Use Bioassay Kits for Biologics

  • Assay design is based on the target biology
  • Measure potency in single- or co-culture models to interrogate appropriate mechanisms of action
  • Contains all components necessary for running the assay
  • High inter- and intra-lot reproducibility (<10% CV)
  • Simple protocol with results in 24 hours or less
  • Seamless method transfer facilitates faster implementation at CMOs, CDMOs, or CROs
  • Amenable to high throughput
 

Bioassay Kit Components

Each bioassay kit is sufficient for at least 1,000 data points and contains the following components:

  • 10 vials of frozen ready-to-assay cells
  • Detection Reagents
  • Cell Plating Reagent
  • Dilution Buffer
  • Control Ligand
  • 10 x 96-well Tissue Culture Treated Plates
 

Bioassay Programs

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

Key Resources

These qualified, off-the-shelf bioassays have been optimized for a particular drug and can reduce assay development timelines by at least 6 months. The simple, easy-to-use assay protocols deliver results fast, enabling implementation into QC lot release and transfer across sites globally.

 
INN Drug Name Qualified With Molecular Mechanism of Action Assay Measures Qualification Data Bioassay Kits Configuration
Aflibercept Eylea® VEGF-Trap VEGFR2 (KDR) Dimerization Request Data New HS Version*
93-0996Y1-00153
93-0996Y1-00154

2-plate
10-plate
Standard
93-0996Y1-00005
93-0996Y1-00006

2-plate
10-plate
Bevacizumab Lucentis® Anti-VEGDA mAb VEGFR2 (KDR) Dimerization Request Data New HS Version*
93-0996Y1-00165
93-0996Y1-00166

2-plate
10-plate
Standard
93-0996Y1-00001
93-0996Y1-00002

2-plate
10-plate
Ranibizumab Avastin® Anti-VEGFA mAb VEGFR2 (KDR) Dimerization Request Data New HS Version*
93-0996Y1-00167
93-0996Y1-00168

2-plate
10-plate
Standard
93-0996Y1-00003
93-0996Y1-00004

2-plate
10-plate
Adalimumab Humira® Anti-TNFα mAb IkB Degradation Request 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 Request 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) Request Data 93-1045B3-00109 2-plate
93-1045B3-00110 10-plate
Anakinra Kineret® IL-1RA (Inhibits IL-1) IL1R/IL1RAP Dimerization Request Data 93-1032Y3-00105 2-plate
93-1032Y3-00106 10-plate
Darbepoetin Alfa Aranesp® EPOR agonist EpoR/EpoR Dimerization Request Data 93-0965Y3-00019 2-plate
93-0965Y3-00020 10-plate
Epoetin Alfa Recombinant hEpo EPOR agonist EpoR/EpoR Dimerization Request Data 93-0965Y3-00017 2-plate
93-0965Y3-00018 10-plate
Exenatide Byetta® GLP1R agonist cAMP Accumulation Request Data 95-0062Y2-00101 2-plate
95-0062Y2-00102 10-plate
Liraglutide Victoza® GLP1R agonist cAMP Accumulation Request Data 95-0062Y2-00099 2-plate
95-0062Y2-00100 10-plate
Insulin Glargine USP Insulin INSRb agonist SH2 Recruitment Request Data 93-0466Y3-00011 2-plate
93-0466Y3-00012 10-plate
Insulin Lispro USP Insulin INSRb agonist SH2 Recruitment Request Data 93-0466Y3-00009 2-plate
93-0466Y3-00010 10-plate
Insulin USP Insulin INSRb agonist SH2 Recruitment Request Data 93-0466Y3-00007 2-plate
93-0466Y3-00008 10-plate
Parathyroid Hormone (PTH) Recombinant hPTH PTHR agonist β-Arrestin Recruitment Request Data 93-0315Y2-00047 2-plate
93-0315Y2-00048 10-plate
Panitumumab ‎Vectibix® Anti-EGFR mAb EGFR/ErbB2 Dimerization Request Data 93-1051Y3-00093 2-plate
93-1051Y3-00094 10-plate
Pertuzumab Perjeta® Anti-HER2-dimerization mAb ErbB2/ErbB3 Dimerization Request Data 93-1042Y3-00095 2-plate
93-1042Y3-00096 10-plate
Teriparatide ‎Forteo® PTHR agonist cAMP Accumulation Request Data 95-0118Y2-00057 2-plate
95-0118Y2-00058 10-plate
Somatotropin, Somatropin Recombinant hGH GHR agonist SH2 Recruitment Request Data 93-0756Y3-00023 2-plate
93-0756Y3-00024 10-plate
Follitropin alfa Gonal-F FSHR agonist cAMP Accumulation Request Data 95-0119Y2-00103 2-plate
95-0119Y2-00104 10-plate
Sargramostim Leukine® Recombinant GM-CSF (CSFR2 agonist) CSF2RB/CSF2R Dimerization Request 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.

Ready-to-use target- or MOA-based bioassays are optimized with a reference ligand. These assays can be readily implemented in development programs for drug candidates (biologics or small molecules) that use the same mechanism of action or the target.

 
Target Molecular Mechanism of Action Assay Measures Bioassay Kits Configuration
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
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

Functional Screening and Cell-Based Potency Assays

Detect Neutralizing Anti-Drug Antibodies (ADAs)

Measure Receptor Dimerization

Optimize Bi-Specific Antibodies

 

Applications of EFC for Biologics Development

 

DiscoverX has exploited the versatile features of Enzyme Fragment Complementation (EFC) technology to create bioassay for a vast number of proteins and applications

 

Receptor Activation

Receptor Internalization

Receptor Dimerization

Receptor Signaling

Receptor Activation Receptor Internalization Receptor Dimerization Receptor Signaling
 

EFC technology can be used to study multiple cell surface receptors and their ligands. By exploiting the natural biology of various receptors, this technology can be used to create simple, cell-based chemiluminescent assays. A. By tagging the receptor and the activation-based intracellular response protein, measure receptor activation, e.g. β-arrestin recruitment to an activated GPCR. B. By tagging the early endosome and the receptor, follow receptor internalization for GPCRs, RTKs, and other receptors. C. By tagging various receptors, monitor the formation of receptor heterodimers and homodimers. D. By tagging cellular pathway proteins, evaluate signaling events downstream of receptor activation including cAMP accumulation, protein translocation, or protein degradation.

 
 

Potency & Stability Testing in QC Lot Release

Eurofins DiscoverX’s high reproducibility bioassays are ideal for determining drug potency and stability using a simple and rapid, mix & read protocol that reflects the drug’s clinical mechanism-of-action. These qualified assays have been optimized for superior bioassay performance to enable a successful regulatory submission package, increasing quality and success of the program.

 

Excellent Assay Reproducibility and Linearity

Excellent Assay Reproducibility and Linearity

 

Bioassays show excellent accuracy and precision. A. PathHunter CHO-K1 CNR1 Bioassay dose response curves for CP 55,940 (a synthetic cannabinoid) demonstrating β-arrestin recruitment in a dose-dependent manner for three days. Table summarizing EC50 values and %RSD for each day; and mean EC50 and %RSD across three days. All three curves show a high degree of overlap for every experiment, thereby demonstrating superior intra-day reproducibility and precision. B. and C. PathHunter Jurkat PD-1 Signaling Bioassay linearity plots showing the average measured relative potency plotted against the expected relative potency for Keytruda and Opdivo, respectively. These assay linearity graphs quantify data generated by two analysts using this bioassay over four days. D. The PD-1 signaling bioassay demonstrates excellent accuracy, precision, and linearity for both Keytruda and Opdivo. Precision values are expressed as a percentage of Relative Standard Deviation (RSD).

 

Highly Specific Responses for MOA-Reflective Bioassays

Highly Specific Responses for MOA-Reflective Bioassays

 

Bioassay measures highly target-specific response to ligands. A. PathHunter Anakinra Bioassay cells expressing the IL-1 receptor (assay quantifies ligand-induced heterodimerization of the IL-1RA and IL-1RAP subunits that comprise the receptor) were treated with various ligands that mediate signaling through the NF-kB pathway, which acts downstream of multiple IL-1 family receptors. Only IL-1β induces a signal in the assay, reflecting the specificity of the assay for IL-1β. B. Two different drugs, Anakinra (which blocks signaling through the IL-1R by blocking IL-1β-mediated dimerization and signaling) and Adalimumab (which binds TNFα, and prevents signaling through the TNFR) were tested in the IL-1R/IL-1RAP bioassay. Only Anakinra blocks IL-1β-mediated dimerization.