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近年来，体内（In Vivo）CAR-T细胞疗法在癌症免疫治疗领域取得了革命性突破，与传统的体外（Ex Vivo）CAR-T疗法不同，In Vivo CAR-T技术通过递送载体将CAR编码基因直接导入患者体内的T细胞，实现免疫细胞的重编程。这一创新疗法不仅简化了治疗流程，还显著降低了生产成本，为癌症患者带来了更便捷、更高效的治疗选择。
Trends Pharmacol Sci. 2024;45(5):406-418.
体内CAR-T细胞疗法工作流程
在In Vivo CAR-T细胞疗法，递送载体的靶向性至关重要。为提升递送效率，通常会用脂质纳米颗粒（Lipid Nanoparticles，LNP）或慢病毒（Lentivirus，LV）等递送载体表面连接靶向抗体，形成抗体偶联递送载体，从而显著增强其对特定细胞（如T细胞、B细胞）的识别与定位能力，实现精准治疗。常见的靶向抗体包括CD3、CD4、CD5、CD7 、CD8等，这些抗体能够有效引导递送载体精准识别目标细胞。递送载体表面偶联抗体的密度和活性，是直接影响该疗法质量、安全性和疗效的关键指标。我们开发了一系列具有确定荧光/蛋白（F/P值）比率的高活性荧光标记蛋白，经严格验证，适用于递送载体表面抗体密度的可靠定量分析。
1e5 of Mouse Anti-CD7 antibody coupled beads (5.5 μm) were stained with different concentration of Alexa Fluor™ 647-Labeled Human CD7 Protein, His Tag (Cat. No. CD7-HA2H4) and negative control protein respectively, AF647 signal was used to evaluate the binding activity (QC tested).
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1e5 of Mouse Anti-CD8 antibody coupled beads (5.5 μm) were stained with different concentration of Alexa Fluor™ 488-Labeled Human CD8 alpha Protein, His Tag (Cat. No. CDA-HA2H6) and negative control protein respectively, AF488 signal was used to evaluate the binding activity (QC tested).
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1e5 of Mouse Anti-CD4 antibody coupled beads (5.5 μm) were stained with different concentration of Alexa Fluor™ 647-Labeled Human CD4 Protein, His Tag (Cat. No. CD4-HA2H8) and negative control protein respectively, AF647 signal was used to evaluate the binding activity (QC tested).
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Alexa Fluor™ 488-Labeled Human CD7 Protein, His Tag (Cat. No. CD7-HA2H9) is stable at 25℃ for 48 hours, equivalent to store at -70℃ for 2 years and freezing and thawing 3 times without performance reduction.
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Binding activity of three different lots of Alexa Fluor™ 488-Labeled Human CD7 Protein, His Tag against Anti-CD7 CAR-293 cells was evaluated by flow cytometry. The result shows very high batch-to-batch consistency.
Immobilized Alexa Fluor™ 647-Labeled Human CD7 Protein, His Tag (Cat. No. CD7-HA2H4) at 1 μg/mL (100 μL/well) can bind Anti-CD7 antibody, Mouse IgG1 with a linear range of 0.05-3 ng/mL (Routinely tested). Labeling with fluorescent dyes did not affect their activity.
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应用案例：验证CD8-LNPs的结合活性与靶向特异性
Alexa Fluor™ 488-Labeled Human CD8 alpha Protein, His Tag (Cat No. CDA-HA2H6) was used to evaluate binding and targeting of CD8-targeted lipid nanoparticles (CD8-LNPs). After, CD8-LNPs were tested to deliver eGFP to immune cells in vitro. Flow cytometry and fluorescence analysis showed high eGFP expression specifically in CD8⁺ cells, confirmed the efficient and selective targeting capability of CD8-LNPs. These results demonstrate the potential of CD8-LNPs for precise delivery of molecular cargo to CD8-expressing cell populations. (Data from Tiva Biosciences).
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