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CD3系列蛋白-双特异性抗体热门靶点

背景
双特异性抗体(bispecific antibody,BsAb)是一类可以同时结合两个不同抗原或者同一抗原上的两个不同抗原表位的抗体分子。双特异性抗体能够发挥两种单抗的协同作用,在临床前研究和临床治疗上可能比混合使用两个单抗更有优势。由于拥有独特的生物学机理,双特异性抗体已经成为抗体药物研发领域的一个重要分支,也是当下制药行业最热门的研发和投资方向之一。根据相关信息统计,目前全球有超过110种双抗处于临床开发阶段,约180种双特异性抗体处于临床前开发阶段,其中,CD3是双抗药物研究的主要靶点[1]
在T细胞表面,CD3分子与TCR以非共价键结合形成TCR/CD3受体复合物。CD3包括四种亚型,分别为CD3δ, CD3ε, CD3γ 和 CD3ζ,其中CD3δ/CD3ε, CD3γ/CD3ε以异源二聚体形式和TCR的α/β链形成TCR-CD3复合物。靶向CD3的抗体药物通常识别形成天然异源二聚体的CD3ε,从而激活T细胞,使其行使肿瘤杀伤功能。然而,在重组表达生产 CD3ε/CD3δ,CD3ε/CD3γ异源二聚体蛋白的过程中,CD3ε与CD3δ或CD3γ极易形成单体比例不均一的复合物,无法保证异源二聚体蛋白的结构和功能,并造成较大批间差异,是双抗药物开发的瓶颈。
ACROBiosystems优化开发了高度均一的高活性CD3δ/ CD3ε, CD3γ/ CD3ε系列产品,经非还原电泳和MALS技术严格确认为1:1异源二聚体,在抗体药物的开发和筛选、抗体药物的鉴定和质控、以及临床血药浓度分析等不同的应用技术平台中均表现出优异的性能,经双特异性临床抗体验证完全符合药物开发要求,可加速抗体药物的开发进程。
产品特点
异源二聚体经MALS验证
经双特异性临床抗体验证
高活性,批间差异小
产品线齐全:his,his&Avi,Fc等tag
验证数据
1:1异源二聚体,经NR电泳和MALS验证

ACROBiosystems开发的高度均一的高活性CD3系列产品,经非还原电泳和MALS技术严格确认为1:1异源二聚体。


Fig.1 Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) on SDS-PAGE under reducing (R) and non-reducing(NR)condition. The purity of the protein was more than 85% and around 35-43 kDa verified by SEC-MALS.

Protocol

高活性,批间差异小

CD3系列产品为严格确认的1:1异源二聚体,更加接近T细胞表面天然蛋白的结构,因此在各种应用场景中表现出高活性。产品高度均一,并采用多种分析技术进行全面严格质控,批间差最小。


Fig.2 Immobilized Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 1 μg/mL (100 μL/well) on Streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate, can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) (Cat. No. CDE-M120a) with a linear range of 0.2-6 ng/mL (QC tested), and batch differences EC50<0.0001 μg/mL.

针对药物开发各应用场景开发

产品适合药物开发的不同阶段的多种应用场景,比如抗体免疫滴度检测,Biopanning抗体筛选,亲和力测定和临床血药浓度分析等。

应用场景1:抗体药物开发和筛选

Fig.3 Immobilized Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.5 μg/well) plate, can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) (Cat. No. CDE-M120a) with a linear range of 0.2-6 ng/mL.

Protocol

Fig.4 Immobilized Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) at 1 μg/mL (100 μL/well) can bind UCHT1 with a linear range of 0.2-6 ng/mL.


Protocol

应用场景2:抗体药物鉴定和质控

Fig.5 Immobilized Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) at 1 μg/mL (100 μL/well) can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) (Cat. No. CDE-M120a) with a linear range of 0.2-6 ng/mL.

Protocol

Fig.6 Immobilized Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (Cat. No. CDD-H52W8) at 5 μg/mL (100 μL/well) can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) (Cat. No. CDE-M120a) with a linear range of 0.4-6 ng/mL.

Protocol

Fig.7 Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) captured on Biotin CAP - Series S sensor Chip can bind Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) with an affinity constant of 22.5 nM as determined in a SPR assay (Biacore T200) .

Protocol

Fig.8 Bispecific T-cell Engager (CD3 X BCMA) immobilized on CM5 Chip can bind Human CD3E&CD3D Heterodimer Protein, His Tag&Tag Free (Cat. No. CDD-H52W1) with an affinity constant of 31.8 nM as determined in a SPR assay (Biacore T200).

Protocol

应用场景3:临床血药浓度分析

Fig.9 Immobilized Human BCMA, His Tag (Cat. No. BCA-H522y) at 2 μg/mL, add increasing concentrations of Bispecific T cell Engager (CD3 X BCMA) in 10% human serum and then add Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 0.2 μg/mL. Detection was performed using HRP-conjugated streptavidin with sensitivity of 15 ng/mL.

Protocol

产品列表
Molecule Cat. No. Species Product Description
CD3E & CD3D CDD-H5258 Human Human CD3E&CD3D Heterodimer Protein, Llama Fc&Llama Fc, low endotoxin
CD3E & CD3D CDD-H52W0 Human Human CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified) HOT
CD3E & CD3D CDD-H52W1 Human Human CD3 epsilon & CD3 delta Heterodimer Protein, His Tag&Tag Free (MALS verified) HOT
CD3E & CD3D CDD-H52W4 Human Human CD3 epsilon & CD3 delta Heterodimer Protein, His Tag&Tag Free
CD3E & CD3D CDD-H52W8 Human Human CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified, Super sensitivity)
CD3E & CD3D CDD-H82F6 Human Biotinylated Human CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His,Avitag™&Fc,Flag,Avitag™ (MALS verified)
CD3E & CD3D CDD-H82W0 Human Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, ultra sensitivity (primary amine labeling)
CD3E & CD3D CDD-H82W6 Human Biotinylated Human CD3 epsilon&CD3 delta Heterodimer Protein, His,Avitag™&Tag Free (MALS verified) HOT
CD3E & CD3D CDD-C5259 Cynomolgus Cynomolgus CD3E&CD3D Heterodimer Protein, Llama Fc&Llama Fc, low endotoxin
CD3E & CD3D CDD-C52W4 Cynomolgus Cynomolgus CD3 epsilon&CD3 delta Heterodimer Protein, His Tag&Flag Tag (MALS verified)
CD3E & CD3D CDD-C52W9 Cynomolgus Cynomolgus CD3 epsilon&CD3 delta Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified)
CD3E & CD3D CDD-C82W6 Cynomolgus Biotinylated Cynomolgus CD3E&CD3D Heterodimer Protein, His,Avitag™&Tag Free (MALS verified)
CD3E & CD3D CDD-M5219 Mouse Mouse CD3 epsilon&CD3 delta Heterodimer Protein, His Tag&Flag Tag (MALS verified)
CD3E & CD3D CDD-M82W5 Mouse Biotinylated Mouse CD3 epsilon&CD3 delta Heterodimer Protein, His,Avitag™&Tag Free
CD3E & CD3G CDG-H52W6 Human Human CD3 epsilon&CD3 gamma Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified, Super sensitivity)
CD3E & CD3G CDG-H52W9 Human Human CD3 epsilon&CD3 gamma Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (MALS verified) HOT
CD3E & CD3G CDG-C5257 Cynomolgus Cynomolgus CD3E&CD3G Heterodimer Protein, Fc Tag&Fc Tag (MALS verified)
CD3E & CD3G CDG-C52W6 Cynomolgus Cynomolgus CD3E&CD3G Heterodimer Protein, His Tag&Tag Free (MALS verified)
CD3 epsilon CDE-H5223 Human Human CD3 epsilon Protein, His Tag
CD3 epsilon CDE-H5256 Human Human CD3 epsilon Protein, Fc,His Tag
CD3 epsilon CDE-H8223 Human Biotinylated Human CD3 epsilon Protein, His Tag, ultra sensitivity (primary amine labeling)
CD3 epsilon CDE-H82E1 Human Biotinylated Human CD3 epsilon Protein, His,Avitag™
CD3 epsilon CDE-C5226 Cynomolgus Cynomolgus CD3 epsilon Protein, His Tag
CD3 epsilon CDE-C5254 Cynomolgus Cynomolgus CD3 epsilon Protein, Fc,His Tag
CD3 epsilon CDE-M5256 Mouse Mouse CD3 epsilon Protein, Fc Tag
CD3 delta CDD-H5224 Human Human CD3 delta Protein, His Tag
CD3 delta CDD-C52H8 Cynomolgus Cynomolgus CD3 delta Protein, His Tag
CD3 epsilon CDE-M120a Mouse Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3), Ultra-low endotoxin
CD3E & CD3D MBS-K003 Human CD3 epsilon&CD3 delta Heterodimer protein-coupled magnetic beads

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