Single B Cell Screening

Overview

Single B cell screening is a powerful technique for isolating and generating antigen-specific monoclonal antibodies (mAbs). Unlike traditional hybridoma technology, this method, often integrated with high-throughput platforms, directly screens primary, naturally matured, individual B cells, and achieves antibody discovery in a time-saving and diversity-protecting manner.Single B cells are advantageous for its simplicity as well, as it requires only a small number of cells and effort needed for obtaining specific mAbs.¹

Biointron's high-throughput single B cell screening platform is capable of screening 2*10^6 plasma B-cells isolated from immunized animals, thus enabling efficient detection of antibody secretion and rapid identification of antibody leads, followed by high through NGS. This platform can accelerate projects such as antibody discovery, vaccine design, and the development of targeted therapies. Especially, it holds significant advantages in discovering antibodies for challenging targets, of rare abundance, or of unique requirement(s).

Highlights

Microfluidic Technology

Microfluidic-based methods can achieve higher throughput and greater flexibility over nanowell and nanovial techniques
This technology has been used in research with single-cell genetics, epigenetics, transcriptomics, proteomics and metabolism

High Efficiency

NGS sequencing & HTP mammalian Production/Validation
Accelerate therapeutic development for a broad range of targets, including challenging transmembrane proteins

Short Timeline

Millions of cells can be screened within 1 week.
By bypassing conventional hybridoma steps with single B cell screening, you can expect to gain 2-3 months.

Speak to A Specialist

Working Flowchart

Mice Immunization

5-10 BALB/c mouse

DNA, Protein, Cells

1-2 months, maintain repertoire diversity

Plasma Cell Enrichment

Isolate plasma B cells from Spleen and Bone Marrow

Single-Cell Droplet Generation and sorting

2E6 plasma B cells are screened

Protein/Cell-based Binding

Single-Cell Sequencing

NGS sequencing

Unique Barcode to make sure the correct pairing

High-Thoughput Expression & Validation

2 weeks for the production

Multiple assays for validation (ELISA, SPR, FACS, Internalization, ADCC, etc.)

Case Study

Case 1: Anti-PD1 Antibody Discovery

Of 62 mAbs expressed, 59 showed good target-binding activity (tested by ELISA).

Of the 59 mAbs, 53 Abs have affinity superior to benchmark antibody (positive control).

Ligand	1:1 binding ka (1/Ms)	kd (1/s)	KD (M)
Ab1	4.26E+05	1.78E-04	4.18E-10
Ab2	4.37E+05	1.86E-04	4.25E-10
Ab3	3.67E+05	1.76E-04	4.80E-10
Ab4	2.27E+05	1.12E-04	4.92E-10
……	……	……	……
Positive control	2.46E+05	1.48E-03	6.01E-09
Ab54	1.90E+05	1.59E-03	8.38E-09
……	……	……	……
Ab59	1.00E+05	2.14E-03	2.14E-08

Diversity analysis indicated excellent CDR protection.

We analyzed the diversity of CDR sequences of the light and heavy chains of 59 Antibodies and showed good diversity.

Epitope binning of all antibodies and found these hits can mainly be divided into 4 epitope groups.

B6924 (Nivolumab biosimilar) and B2014 (Pembrolizumab biosimilar) are positive controls. The result showed that we have identified many antibodies binding to distinct epitopes on PD-1 compared to controls.

Case 2: Anti-TAA Antibody Discovery

Of 70 mAbs expressed, 45 showed good target-binding activity (tested by ELISA).

Six molecules (samples) evaluated in cell-based blocking assay showed better blocking activity than positive control antibody.

No.	IC50	maximum inhibition rate
Sample1	6.62	90.96%
Sample2	5.91	90.14%
Sample3	84.88	87.33%
Sample4	11.18	85.69%
Sample5	32.21	87.77%
Sample6	52.85	80.63%
Positive Control	329.2	/

"Every single B cell screening holds the potential for groundbreaking discoveries, and I am honored to be part of a team that is pushing the boundaries of what's possible. I am genuinely excited to contribute to the future of scientific discovery."

Rui Zhou

Single B Cell Screening Team

FAQs

What method does Biointron use for Single B Cell Screening?

The method that Biointron uses is a microfluidic-based platform. This allows us to analyze individual B cells and their antibody repertoires with high throughput and precision. It involves miniaturized channels and chambers on a microchip, where single B cells are captured and isolated. We can perform various analyses on each cell, including gene sequencing, functional assays, and phenotypic characterization.
Do other methods exist?
There are several other methods for Single B Cell Screening, such as:
- Fluorescence-Activated Cell Sorting (FACS): B cells expressing specific cell surface markers or antigen-binding receptors can be isolated and analyzed at a single-cell level using FACS to study the diversity of B cell populations.
- Single-Cell PCR and RT-PCR: These can be applied to individual B cells to amplify and analyze the genetic information of their antibody genes, in order to determine nucleotide sequences of antibody variable regions.
- Single-Cell RNA Sequencing (scRNA-seq): scRNA-seq profiles the transcriptome of individual cells, provides a comprehensive view of gene expression patterns.²
Why use Single B Cell screening for antibody discovery?

The project lead time is much faster than hybridoma or phage display methods, being 3 months rather than 6 months and 5 months, respectively. The library size is larger than hybridoma methods, and the Single B Cell platform uses natural binding and a highly automatic process control.

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