阿拉玛蓝色测试优化以最大限度地减少药物干扰和测试间可行性
Mina N Dinh1, Masahiro Hitomi1, Zahraa A Al-Turaihi1
1Department of Translational Hematology & Oncology Research, Cleveland Clinic, Cleveland, OH, United States of America.
MethodsX
|November 18, 2024
概括
一种经过修改的Alamar Blue (AB) 试验可以防止因EGFR抑制剂引起的错误高细胞活力读数. 这种具有成本效益,非破坏性的方法提高了癌症研究的准确性,使得可靠的剂量反应曲线分析成为可能.
科学领域:
- 生物化学 生物化学
- 细胞生物学 细胞生物学
- 药理学 药理学是指药理学的学科.
背景情况:
- 阿拉玛蓝 (AB) 是一种受欢迎的,具有成本效益的,用于细胞活力测试的非破坏性试剂.
- 之前的研究已经注意到,当使用AB与其他测定方法 (如Cell-Titer Glo) 相比,特别是使用EGFR抑制剂时,剂量反应曲线存在差异.
- 调查奥西默提尼布对非小细胞肺癌 (NSCLC) 细胞系PC-9的影响揭示了AB测定剂量反应曲线中意想不到的右转.
研究的目的:
- 描述一种修改后的Alamar Blue测试方法,以克服测试干扰并实现准确的剂量反应曲线.
- 优化Alamar Blue测试协议,以提高细胞活力和药物反应研究的可靠性.
- 建立Alamar Blue作为一种适合细胞生长和药物毒性恢复的纵向监测方法.
主要方法:
- 修改后的方法包括在Alamar Blue添加药物之前去除含药介质,以防止干扰.
- 通过结合光校准标准来调整度仪的灵敏度,从而最大限度地减少了板块对板块的变化.
- 通过比较剂量反应曲线与来自Cell Titer Glo试验的曲线来验证修改后的试验.
主要成果:
- 移除含有药物的介质成功地消除了错误的升高读数,产生与Cell Titer Glo试验相比的剂量反应曲线.
- 添加光校准标准有效地减少了板对板的变化,确保了一致的测试灵敏度.
- 经过修改的Alamar Blue测试显示,在评估药物对癌细胞系的影响时,其准确性与Cell Titer Glo相美.
结论:
- 修改后的Alamar Blue试验提供了准确可靠的细胞活力数据,与 Cell Titer Glo.等既定方法相比.
- 这种优化的协议解决了以前的局限性,使得Alamar Blue成为癌症药物疗效研究的强大工具.
- 阿拉玛蓝色试验可用于细胞健康和细胞毒性治疗后恢复的持续纵向监测.
更多相关视频
08:23Design and Development of Aptamer–Gold Nanoparticle Based Colorimetric Assays for In-the-field Applications
Published on: June 23, 2016
12.2K
11:03Use of Capillary Electrophoresis Immunoassay to Search for Potential Biomarkers of Amyotrophic Lateral Sclerosis in Human Platelets
Published on: February 10, 2020
7.2K
相关概念视频
01:15Bioavailability Study Design: Healthy Subjects Versus Patients
Bioavailability studies are essential for evaluating a drug's therapeutic efficacy and understanding its absorption patterns under various physiological conditions. Conducting such studies on target patient populations provides more relevant data by simulating real-world disease states. However, practical challenges often necessitate the use of young, healthy adult volunteers as study subjects.Patients may exhibit altered drug absorption patterns due to the effects of the disease itself,...
01:26Therapeutic Drug Monitoring: Drug Analysis Methods
Therapeutic Drug Monitoring (TDM) is a clinical practice that measures specific drug levels in a patient's blood or body tissues to tailor drug therapy effectively. This monitoring is critical for managing drugs with narrow therapeutic indices like digoxin and phenytoin, ensuring they are both safe and effective. For instance, monitoring theophylline levels in asthma patients involves precision and sensitivity to adjust doses according to individual responses to therapy, ensuring efficacy and...
01:22Effect of Hepatic Disease on Pharmacokinetics: Pathophysiologic Assessment and Liver Function Test
In clinical practice, the direct measurement of hepatic blood flow to evaluate liver function presents significant challenges due to the intricate and specialized nature of the necessary techniques. Consequently, healthcare professionals often rely on empirical estimates derived from thorough patient examinations and liver function tests to gauge liver health. Among the tools at their disposal, the Child–Pugh and MELD scoring systems stand out for their ability to categorize and assess the...
01:23Toxicity Testing in Animals
Toxicity tests in animals are grounded on two main assumptions: first, the effects observed in laboratory animals can be extrapolated to humans, especially when adjusted for body surface area; second, high-dose exposure in animals is essential to identify potential human hazards from lower doses. This is based on the quantal dose-response concept, which faces the challenge of extrapolating results from relatively few test animals to much larger human populations. For example, a 0.01% incidence...
01:28Antimicrobial Effectiveness
The effectiveness of antimicrobial agents depends on various factors influencing their ability to eliminate microbial populations. Larger microbial populations require more time for complete eradication, emphasizing the importance of population size analysis when evaluating antimicrobial efficacy.Microbial resistance to antimicrobial agents varies significantly. Highly resilient microorganisms include endospores, gram-negative bacteria, and non-enveloped viruses, while prions are exceptionally...
01:28Methods of Medium Optimization
Optimizing growth media enhances microbial proliferation and maximizes product yield. Statistical experimental design methodologies provide structured and reproducible approaches, offering progressively higher levels of robustness and efficiency.The One-Factor-at-a-Time (OFAT) MethodThe One-Factor-at-a-Time (OFAT) method involves adjusting a single variable while keeping all others constant. However, it cannot detect interactions between variables, often leading to suboptimal outcomes when...