About the Hypercell^TM System

America Diagnostics' Hypercell® System is a groundbreaking high-throughput single-cell sorting platform that accelerates antibody discovery and cloning. By combining automation, precision, and speed, the Hypercell® System significantly outperforms traditional methods such as phage display, yeast display, and hybridoma technologies.

Key Features

• High Efficiency: Screens hundreds of thousands of secretory cells within hours.
• Flexibility: Compatible with multiple cell and assay types.
• Small Footprint: Bench-top design fits in standard biosafety cabinets.
• Integrated System: Combines flow sorting, detection, and control for seamless operation.
• User-Friendly: Minimal hands-on time with automated processes.

Why Choose Hypercell^TM?

Compared to traditional technologies, Hypercell® offers:

With the Hypercell^TM System, researchers can achieve higher antibody affinity and diversity in a fraction of the time, making it the ultimate choice for modern antibody discovery.

System Workflow

The Hypercell^TM System workflow is streamlined to deliver results in hours, not weeks !

Discover cutting-edge antibody screening with our AmerIDx® system, featuring the Hypercell^TM technology. This efficient, step-by-step process is designed to optimize antibody discovery for biomedical research and therapeutic applications. Here's an overview of the workflow:

1. Plasma B cells enrichment (~1-2h): Enrich plasma B cells from a mouse.
2. Encaps single cell & Hypercell^TM reagents Incubation (~1-1.5h): Encapsulate single cells with Hypercell® reagents and incubate.
3. Hypercell® sorting (~1-2h): Sort the cells using the Hypercell^TM system.
4. Target cells recovery (~0.5h): Recover the target cells.
5. Single cell RT-PCR & VH/VL sequencing (~5 days): Perform single-cell RT-PCR and VH/VL sequencing.
6. AbFinder™ bioinformatic analysis (<1 day): Conduct bioinformatic analysis with AbFinder™.
7. Ab validation (~14-21 days): Complete antibody validation.

   Unlock the potential of advanced antibody discovery with our streamlined workflow, ensuring precise and reliable results.

   

   Fig 1. Hypercell^TM workflow for antibody screening, from immunized animal to purified antibody.
   Our process includes single-cell sorting, target cell recovery, and sequence analysis with AbFinder™ software, all designed for unparalleled efficiency.

Case Studies

Explore the real-world performance of the Hypercell^TM System:

Case Study 1: B2M (beta2 macroglobin ) Antibody Discovery in Plasma B Cells

To identify high-affinity binders from plasma B cells and evaluate their sequence diversity and binding properties using the Hypercell^TM System and AbFinder^TM software.

Workflow and Key Steps

1. Plasma B Cell Enrichment:
• B cells were enriched from immunized animals.
• The enriched cells were incubated with Hypercell® reagents to prepare them for single-cell sorting.
2. Single-Cell Sorting and Analysis:
• Using the Hypercell^TM System, individual plasma B cells secreting antibodies of interest were encapsulated in droplets and sorted based on secretion rates and binding properties.
• The system identified 42 binders in Mouse #1.
3. VH and VL Sequence Analysis:
• Following single-cell sorting, VH and VL regions of antibody genes were sequenced using RT-PCR.
• The sequence data was analyzed with the AbFinder™ software, which generated:
• VH-VL Circos Plots to illustrate sequence relationships.
• Phylogenetic Trees to show diversity among antibody variants.
4. Binding Affinity Assessment:
• ELISA and Bio-Layer Interferometry (BLI) were used to validate binding properties.
• Key binding parameters such as association rate (kon), dissociation rate (koff), and affinity constant (KD) were calculated.

Results and Key Metrics

1. Binder Identification:
• 42 binders were selected, with 67.7% binder ratio based on sequence analysis.
• When combined with structural docking analysis, 17 binders were confirmed as high-affinity candidates, with a success rate of 77.3%.


Fig 2. Key numbers in Case study 1

2. Sequence Diversity:
• Analysis of VH and VL sequences showed a broad range of somatic hypermutations.
• CDR3 Length Distribution highlighted significant variability, a hallmark of a diverse antibody repertoire.
3. Binding Validation (BLI Data):
• Multiple antibodies demonstrated strong binding affinities:
• Ab196: KD = 2.17 × 10⁻¹⁰ M, indicating exceptionally high affinity.
• Ab507: KD = 7.19 × 10⁻¹⁰ M.
• Ab130: KD = 4.69 × 10⁻⁹ M.
• These results validated the Hypercell® System's ability to identify binders with high specificity and affinity.
4. ELISA Results:
• ELISA confirmed the functional binding of mAbs to the target antigen.
• Positive control wells demonstrated robust activity, matching BLI kinetics.

Fig 3. ELISA results in in Case study 1

Highlighted Features of the Hypercell® System in This Case

1. High-Throughput Screening:
• The system screened thousands of plasma B cells rapidly, identifying binders within hours.
2. Detailed Sequence Analysis:
• AbFinder™ software enabled efficient VH/VL sequence analysis, revealing high levels of diversity and guiding lead selection.
3. Precision Binding Assessment:
• Identified antibodies showed strong binding affinities and specificities, confirmed through both BLI and ELISA.
4. Rapid Workflow:
• From cell sorting to sequence analysis and binder validation, the entire process was completed significantly faster than traditional methods.

Conclusion

In this case, the Hypercell^TM System enabled rapid identification and characterization of high-affinity antibodies, significantly accelerating the antibody discovery process. The system’s integration with AbFinder™ software provided seamless analysis of sequence diversity and functionality, highlighting its value as a next-generation solution for therapeutic antibody development.

Hypercell® identified high-affinity antibodies in one day, significantly faster than traditional methods. The AbFinder™ software facilitated sequence analysis and lead selection.

Case Study 2: Stability Test of Antibodies

To evaluate the effectiveness of the Hypercell® System in discovering high-affinity antibodies against HBsAg (Hepatitis B surface antigen), a key diagnostic and therapeutic target for Hepatitis B Virus (HBV).

Workflow and Key Steps

1. Plasma B Cell Enrichment:
• Plasma B cells were enriched from immunized mice (#1 and #2).
• Enriched cells were subjected to Hypercell® sorting to identify antibody-secreting cells targeting HBsAg.
2. Single-Cell Sorting and Analysis:
• The Hypercell® System encapsulated and analyzed individual plasma B cells.
• Secretory activity and binding to HBsAg were used to select high-affinity candidates.
3. Sequence Analysis with AbFinder™:
• VH and VL regions of selected antibody genes were sequenced and analyzed.
• Phylogenetic trees were generated to illustrate antibody sequence diversity and relationships.
4. Binder Validation:
• ELISA assays confirmed binding activity of the selected antibodies.
• Binding kinetics (affinity and specificity) were measured using BLI (Bio-Layer Interferometry).

Results and Key Metrics

1. Binder Selection:
• A total of 20 binders were identified from the pooled analysis of Mouse #1 and Mouse #2, with a binder ratio of 58.8%.
• Individual results:
• Mouse #1: Binder ratio of 67.7%.
• Mouse #2: Binder ratio of 76.4%.


Fig 4. ELISA results in in Case study 2

2. Binding Affinity (BLI Data):
• Key binders showed strong binding kinetics:
• Ab36: KD = 1.63 × 10⁻¹² M, indicating exceptionally high affinity.
• Ab140: KD = 4.02 × 10⁻¹⁰ M.
• Ab2006: KD = 1.37 × 10⁻¹¹ M.


Fig 5. BLI Kinetics data in Case study 2

3. Sequence Diversity:
• Phylogenetic analysis revealed high diversity in the antibody sequences, with significant variations in VH and VL regions.
• High sequence diversity supports the identification of unique, high-affinity antibodies.
4. ELISA Validation:
• ELISA results confirmed strong binding activity for all selected antibodies, with low EC50 values (e.g., Ab36: EC50 = 0.015 nM).

Highlighted Features of the Hypercell^TM System in This Case

1. Efficient Screening:
• The Hypercell^TM System rapidly sorted and analyzed plasma B cells, identifying binders in a fraction of the time required by traditional methods.
2. High Binding Affinity:
• Identified antibodies exhibited exceptionally low KD values, reflecting high binding strength to HBsAg.
3. Sequence Analysis with AbFinder^TM:
• Detailed VH/VL analysis provided insights into antibody diversity and guided the selection of promising candidates.
4. Reduced Time to Results:
• The entire process, from cell sorting to binder validation, was completed in days, compared to weeks or months with conventional workflows.

Conclusion

Case Study 2 demonstrates the Hypercell^TM System's ability to efficiently identify high-affinity antibodies against HBsAg. The system's integration of single-cell sorting, high-throughput analysis, and AbFinder™ bioinformatics enabled rapid and precise antibody discovery, making it a powerful tool for therapeutic and diagnostic antibody development.

Demonstrated superior performance in stability tests for antibodies generated using the Hypercell^TM platform compared to conventional approaches.