Comparation Of Two Immunomagnetic Nanobeads In PIG-A Gene Mutation Assay In Vitro Volume 5 - Issue 5

Yi Zhang, Jiaqi Lv, Xiao Chen, Yulu Li, Haitao Yang, Qing Miao, Baolier Wuhan, Jingwei Xiao* and Binli*

• National Institute of Occupational Health and Poison Control, CHINA CDC

Received:September 06, 2021;   Published:September 21, 2021

Corresponding author: Jingwei Xiao, Binli, National Institute of Occupational Health and Poison Control, CHINA CDC

DOI: 10.32474/SJPBS.2021.05.000223

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Abstract

Objective: The enrichment ability was compared between two kinds of immunomagnetic nanobeads in phosphatidylinositol glycans A (PIG-A) gene mutation assay.
Methods: Glycosylphosphatidylinositol(GPI)(+) TK6 cells was enriched and GPI (-) TK6 cells were removed by both PE and Biotin immunomagnetic nanobeads. Flow cytometry was used to detect the purification effect. PIG-A gene mutation assay was performed on purified TK6cells with typical positive Ethyl methane Sulfonate (EMS).
Results: After once purification, PE immunomagnetic nanobeads could reduce the GPI(-)background value from 44.90±1.63% to 0.27±0.12%, while the value of Biotin immunomagnetic nanobeads was 4.27±0.76%.Compared with the control group ,the mutation frequency of PIG-A gene increased significantly at 50μg/mL on the 12th day, at 100μg/mL and 200μg/mL on the 4th, 8th and 12th day.
Conclusions: PE immunomagnetic nanobeadswere more suitable than Biotin immunomagnetic nanobeads for TK6 cells purification of PIG-A gene mutation assay in vitro.

Keywords: PE Immunomagnetic Nanobeads; Biotin Immunomagnetic Nanobeads; PIG-A Gene Mutation; EM; Flow Cytometry

Abbreviations: EMS:Ethylmethane Sulfonate; PIG-A: phosphatidylinositol glycans A; GPI:glycosylphosphatidylinositol; FBS:Fetal Bovine Serum; PBS:Phosphate Buffer Solution.

Abstract| Introduction| Materials and Methods| Ems Treatment| Flow Cytometry Analysis| Statistical Analysis| Results| Discussion| Acknowledgement| Disclosure of Interest| References|