シングルB細胞スクリーニング(Single B Cell Screening)は、特異的なモノクローナル抗体（mAb）を作製するための強力な技術です。この方法は、ハイスループットプラットフォームと統合され、抗体分泌の効率的な検出とリードクローンの迅速な同定を可能にし、抗体の開発、ワクチン設計、および標的治療の開発を加速します。シングルB細胞は、わずかな細胞数で特異的なmAbを迅速に得るための高い効率を維持しつつ、その単純さが利点とされています。
Biointronは、抗体作製を加速するためのハイスループット・シングルB細胞スクリーニング(Single B Cell Screening)・プラットフォームを構築しています。このプラットフォームを使用すると、免疫動物から採取した2*106個のプラズマB細胞を1回のスクリーニングで処理でき、特異的な抗原に対する抗体を分泌する細胞を単離することも可能です。これにより、非常に希少な抗体や、ユニークな性質を持つ潜在的な薬物を検出することができるようになりました。
5〜10匹のBALB/cマウス
DNA、タンパク質、細胞
レパートリーの多様性を維持しながら1〜2か月
脾臓と骨髄からプラズマB細胞を単離
2E6プラズマB細胞をスクリーニング
タンパク質/細胞ベースの結合
NGSシーケンシング
ユニークなバーコードで正しいペアリングを確認
製作期間：2週間
バリデーションのための複数のアッセイ（ELISA、SPR、FACS、インターナリゼーション、ADCCなど）
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.
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 | / |
Single B cell screening is a technique for isolating and generating antigen-specific monoclonal antibodies by directly screening individual B cells. This method preserves the natural pairing of heavy and light chains, leading to the discovery of antibodies with high specificity and affinity.
Biointron's platform utilizes microfluidic technology to screen up to 2 million plasma B cells from immunized animals. The process includes isolating plasma B cells, generating single-cell droplets, sorting based on antigen binding, sequencing, and high-throughput expression and validation.
Microfluidic-based methods achieve higher throughput and greater flexibility compared to traditional techniques. They allow for efficient handling of small volumes, precise control over experimental conditions, and integration of multiple steps into a single platform, enhancing the efficiency and speed of antibody discovery.
The timeline varies based on project complexity. Biointron's platform can screen millions of cells within one week, and by bypassing conventional hybridoma steps, the process can save 2–3 months, leading to a shorter overall project duration.
The platform is versatile and can target a broad range of antigens, including challenging transmembrane proteins, proteins, peptides, and small molecules. This flexibility makes it suitable for various antibody discovery projects.
Biointron employs multiple validation assays, such as ELISA, SPR, FACS, internalization studies, and ADCC assays, to confirm the functionality and specificity of the antibodies. Additionally, next-generation sequencing (NGS) is used to ensure accurate pairing of heavy and light chains.
Yes, with high-capacity automated systems and a dedicated team, Biointron can manage multiple antibody discovery projects concurrently without compromising quality.
Clients receive detailed reports, including sequence information, binding affinity data, functional assay results, and purified antibody samples for further testing.
Yes, all intellectual property rights for antibodies discovered under client-specific projects are fully transferred to the client.
To start, contact Biointron through the website’s inquiry form or email. A consultation will be arranged to discuss project requirements and provide a tailored proposal.
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