使用质谱免疫分析法通过液相色谱-串联质谱法对猴血清中的人IgG1进行定量分析。
Quantitative analysis of hIgG1 in monkey serum by LC-MS/MS using mass spectrometric immunoassay.
作者信息
Lanshoeft Christian, Heudi Olivier, Cianférani Sarah, Warren Andrew P, Picard Franck, Kretz Olivier
机构信息
Novartis Institutes for Biomedical Research, Drug Metabolism & Pharmacokinetics, Novartis Campus, Fabrikstrasse 14, 4056 Basel, Switzerland.
Laboratoire de Spectrométrie de Masse BioOrganique, Institut Pluridisciplinaire Hubert Curien, CNRS-Université de Strasbourg, UMR 7178, 25 rue Becquerel, 67087 Strasbourg, France.
出版信息
Bioanalysis. 2016 May;8(10):1035-49. doi: 10.4155/bio.16.32. Epub 2016 Apr 20.
AIM
A sensitive generic LC-MS/MS method for hIgG1 quantification in cynomolgus monkey serum using mass spectrometric immunoassay disposable automation research tips (MSIA-D.A.R.T.'S™) is reported.
RESULTS
The hIgG1 was captured with a biotinylated mouse anti-hIgG antibody (50.0 µg/ml) targeting the fragment crystallizable (Fc) region. Elution from the streptavidin-coated MSIA-D.A.R.T.'s was conducted with 0.4% trifluoroacetic acid in water. The method was selective and linear from 10.0 to 1000 ng/ml using 100 µl of serum. The method was evaluated regarding accuracy, precision, carry-over, dilution, auto-sampler stability and applied for the determination of hIgG1 concentration in monkey serum after intravitreal administration.
CONCLUSION
The present assay is suitable for quantitative analysis of hIgG1-based therapeutic proteins in monkey serum at low levels.
目的
报道一种使用质谱免疫分析一次性自动化研究提示（MSIA-D.A.R.T.'S™）在食蟹猴血清中定量检测人IgG1的灵敏通用液相色谱-串联质谱（LC-MS/MS）方法。
结果
使用靶向可结晶片段（Fc）区域的生物素化小鼠抗人IgG抗体（50.0μg/ml）捕获人IgG1。用含0.4%三氟乙酸的水溶液从包被抗生物素蛋白链菌素的MSIA-D.A.R.T.'S上洗脱。该方法具有选择性，使用100μl血清时在10.0至1000ng/ml范围内呈线性。对该方法的准确性、精密度、残留、稀释、自动进样器稳定性进行了评估，并应用于玻璃体内给药后猴血清中人IgG1浓度的测定。
结论
本测定法适用于低水平猴血清中基于人IgG1的治疗性蛋白质的定量分析。
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