アフィニティ・マチュレーション(Affinity Maturation)は、まだ親和性成熟といいます。抗体と標的抗原への結合強度を向上させるために使用されます。抗体最適化方法の1つです。 これは、哺乳動物のB細胞における体内でのソマティックハイパーミューテーションとクローン選択によって自然に行われますが、治療目的のために実験室内でのミュータジェネシスと選択によっても行われることがあります。
FCMES-AM (Full Coverage Mammalian Expression System for Affinity Maturation) は、Biointronの特許プラットフォームです。CDR領域内の各アミノ酸は、システインとメチオニンを除く17種類のアミノ酸に均等な割合で変異させます。クローンは哺乳類系で発現され、最初のテストとしてELISA結合アッセイが実施されます。これに続いて、さらなる親和性測定のためにSPR (surface plasmon resonance)またはFACS (fluorescence-activated cell sorting) が行われます。
Biointronの専門チームにお気軽にお問い合わせください。私たちのプラットフォームが、より優れた性能のためにあなたの抗体の親和性を向上させる方法を探るお手伝いをいたします。
私たちのプラットフォームにより、抗体の親和性を向上させ、優れた性能の抗体を提供します。お気軽にお問い合わせください。
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ホット・スポットの特定 |
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抗体発現とアフィニティー検出 |
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The parental antibody is VHH, after affinity maturation, off of the VHH improves and the affinity increases 23-fold, 19-fold and 6-fold separately.
The parental antibody is mouse IgG, after affinity maturation, the affinity increases 120-fold, 48-fold and 46-fold separately.
Affinity maturation is widely used to improve antibody activity through mutagenesis and selection for downstream applications such as therapeutics and diagnostics. This is often necessary to ensure the affinity towards the target and that the therapeutic potential of antibodies is reached. Optimization can also involve adjusting functionality, cross-reactivity, stability, immunogenicity, and any chemical liabilities.1
In vivo
When a host is repeatedly exposed to the same antigen, the antibodies produced by their immune response will have increasingly greater affinity, avidity, and anti-pathogen activity. This occurs due to somatic hypermutation in the variable CDRs (complementarity-determining regions) of immunoglobulin genes in B cells, and takes place in germinal centers (structures within secondary lymphoid tissues). Clonal selection takes place when follicular dendritic cells of the germinal centers present antigens to the B cells, with only the most competitive B cells surviving to stably conjugate with follicular B helper T cells. After several rounds of selection, high-affinity antibodies will be produced.2
In vitro
Similar to the in vivo process, in vitro affinity maturation is used to optimize antibodies, antibody fragments, or other peptides by diversifying the antibody base sequence and isolating higher-affinity binders.
Although antibodies can be generated against countless distinct epitopes, your genome does not have the capacity to hold all the information. Thus, B-cells employ somatic hypermutation, which involves mutations of the genes which encode the antigen-binding region in direct response to the antigen stimulus. Through successive generations and exposure, only B cells producing antibodies that recognize the antigen with high affinity will survive.3