The Biodesign Institute at Arizona State University, Tempe, AZ 85287 USA.
The Biodesign Institute at Arizona State University, Tempe, AZ 85287 USA ; Department of Chemistry & Biochemistry at Arizona State University, Tempe, AZ 85287 USA.
Proteome Sci. 2014 Oct 14;12(1):52. doi: 10.1186/s12953-014-0052-3. eCollection 2014.
The cytokine MIF (Macrophage Migration Inhibitory Factor) has diverse physiological roles and is present at elevated concentrations in numerous disease states. However, its molecular heterogeneity has not been previously investigated in biological samples. Mass Spectrometric Immunoassay (MSIA) may help elucidate MIF post-translational modifications existing in vivo and provide additional clarity regarding its relationship to diverse pathologies.
In this work, we have developed and validated a fully quantitative MSIA assay for MIF, and used it in the discovery and quantification of different proteoforms of MIF in serum samples, including cysteinylated and glycated MIF. The MSIA assay had a linear range of 1.56-50 ng/mL, and exhibited good precision, linearity, and recovery characteristics. The new assay was applied to a small cohort of human serum samples, and benchmarked against an MIF ELISA assay.
The quantitative MIF MSIA assay provides a sensitive, precise and high throughput method to delineate and quantify MIF proteoforms in biological samples.
细胞因子 MIF（巨噬细胞移动抑制因子）具有多种生理作用，在许多疾病状态下其浓度升高。然而，其在生物样本中的分子异质性尚未得到研究。质谱免疫分析（MSIA）可能有助于阐明体内存在的 MIF 翻译后修饰，并为其与多种病理的关系提供更多的明确信息。
在这项工作中，我们开发并验证了一种用于 MIF 的完全定量 MSIA 测定法，并将其用于在血清样本中发现和定量不同形式的 MIF，包括半胱氨酸化和糖化 MIF。MSIA 测定法的线性范围为 1.56-50ng/mL，具有良好的精密度、线性和回收率特征。新测定法应用于一小部分人血清样本，并与 MIF ELISA 测定法进行了基准测试。
定量 MIF MSIA 测定法提供了一种灵敏、精确和高通量的方法，可用于描绘和定量生物样本中的 MIF 蛋白形式。
