嵌合抗体会表达在细胞表面，然后用流式检测吗？

Veblen

请教老师，在这篇文章中提到嵌合抗体的检测，但是又提到了表面染色来检测比较MFI,我看了很久没看懂。

例如：Fig. 8B中 Although no significant difference was observed in the surface staining of the secreted chimeric antibody on day 4, MFI was significantly increased on day 24 (Fig. 8B; Fig. S14B). MFI increase was 2.14 ±0.10-fold for ROSE_Blimp1α and 2.17 ± 0.03-fold for ROSE_Blimp1β, compared to their respective uninduced states.
请问这里的嵌合抗体是如何用流式检测的？文章中好像提到的是表面染色，为什么嵌合抗体可以表面染色就可以检测？
翻来覆去看了好几遍，一直无法理解，所以诚心请教一下老师，感谢，感谢。

附近是全文，再次感谢。

niwanmao

ROSE_Blimp1α和ROSE_Blimp1β本身是转录因子，主要在细胞核内发挥作用，但它们对嵌合抗体基因表达的影响可以通过检测细胞表面或分泌的嵌合效应分子EGFP和TagRFP657来间接评估。通过测量的表达水平变化，可以推断Blimp1的活性和功能状态。


原文：
For flow cytometry analysis, adherent cells were resuspended in PBS +1% FBS, and suspension cells were prepared in culture media. FACSCalibur (Becton Dickinson, Franklin Lakes, NJ, USA) was used as previously described (Shin and Lee, 2020b) for SSI-based CMV-Tet-On3G experiments. Unless otherwise specified, Novocyte 2000R (Agilent, Santa Clara, CA, USA) was used for flow cytometry analysis. A 488 nm blue laser and a 530/30 filter were used to detect EGFP and AF488, and a 640 nm red laser and 675/30 filter was used to detect TagRFP657. Raw data were analyzed using NovoExpress software (v.1.6.2) and visualized using R software (version 4.1.0).

Veblen

niwanmao 发表于 2024-11-23 11:05
ROSE_Blimp1α和ROSE_Blimp1β本身是转录因子，主要在细胞核内发挥作用，但它们对嵌合抗体基因表达的影响可 ...

非常感谢老师，恍然大悟；
Suddenly be enlightened, thank you.