Single-Cell Cloning Platform (Centre of Excellence)

Head: HUO Jianxin, Ph.D.

Email: huo_jianxin@immunol.a-star.edu.sg

Single-Cell Cloning Platform advances antibody discovery, immune cell profiling and cell line development using the Beacon Optofluidic System

Introduction

A*STAR SIgN and Bruker Cellular Analysis proudly announces their latest Centre of Excellence (CoE), the Single Cell Cloning Platform, which utilizes the Beacon Optofluidic System, an innovative technology that seamlessly combine Optoelectronic Positioning (OEP) and microfluidics to offer rapid and precise cell sorting, single-cell functional analysis, and real-time monitoring of cellular interactions and responses. Highly efficient and high-throughput, the platform facilitates advanced single-cell research and supports biopharmaceutical applications.

The Beacon OptoFluidic System supports a range of advanced applications, including:

• Antibody Discovery: Facilitates the identification of high-affinity antibodies across various species, including mouse, human, and rabbit.
• Immune Cell Profiling: Enables detailed analysis of immune cell activation, cytokine secretion, and target cell apoptosis, providing valuable insights into immune responses.
• Cell Line Development: Streamlines the screening and isolation of high-producing cell clones, enhancing efficiency in developing optimal cell lines for specific applications.
• Customized Single-Cell Applications: Offers tailored solutions to meet unique research needs, allowing for flexible exploration in diverse fields of study.
  

Technologies and Approaches

Figure 1: Beacon System (left); Figure 2: OptoSelect Chips (right)

At the core of the Beacon Optofluidic System is the OptoSelect chip, containing thousands of NanoPen chambers. It employs OEP, which uses light for the large-scale manipulation of cells, directing single cells into NanoPens with the additional assistance of AI. (Figure 3)

Antibody Discovery and Immune Cell Profiling

The Beacon Optofluidic System revolutionizes antibody discovery by actively monitoring cell-antigen interactions, enabling the detection of promising candidates for high-affinity antibody-producing plasma cells. (Figure 4). 

This process involves:

• Real-time IgG Production: Continuous tracking of IgG levels allows for immediate assessment of antibody response. (Figure 5)
  

Figure 5: Antibody Discovery - Assay with IgG Beads

• Specific Antibody Generation Assays: Targeted assays provide insights into the generation of specific antibodies. (Figure 6)

Figure 6: Antibody Discovery - Assay with Antigen-Presenting Beads

Once the promising plasma B cells are identified, they can be exported for comprehensive analysis and characterization. This ensures a streamlined pathway from discovery to detailed study and enhances the efficiency and accuracy of antibody development efforts.

The Beacon also facilitates detailed profiling of T and NK cells by culturing them as single cells within a microfluidic environment, where they can be activated by targeted cells.

Key features of this system include:

• Real-time Imaging and Biosensors: These technologies enable precise measurement of cellular responses, including:
• Cytokine Production: Monitoring the release of cytokines helps assess immune activation. (Figure 7)
• T/NK Cell Activation: Evaluating the activation status of these cells is crucial for understanding immune responses. (Figure 8)
• Target Cell Apoptosis: Tracking the apoptosis of target cells provides insights into the effectiveness of immune responses. (Figure 8)

Figure 7: T-Cell Profiling - Cytokine Secretion

Figure 8: T-Cell Profiling - Cancer Cell Killing

Cell Line Development

The Beacon® OptoFluidic System accelerates cell line development by providing several key features:

• Single-Cell Isolation: This capability allows for the precise selection of individual cells, ensuring the generation of homogeneous cell populations.
• Controlled Culturing: Cells are cultured in a carefully managed environment, promoting optimal growth conditions (see Figure 9).
• Clonal Selection: The system facilitates the identification of desirable clones based on specific criteria, enhancing the efficiency of the selection process.
• High-Throughput Screening: Large numbers of cell lines can be evaluated simultaneously, significantly speeding up the screening process.

With real-time monitoring and advanced data analysis, the system streamlines the identification of optimal cell lines tailored for specific applications. Once selected, these cells can be efficiently exported for large-scale expansion (see Figure 10), paving the way for applications in research, therapeutics, and biopharmaceutical production.