■Ravulizumab(RAVUL)是一种新的补体抑制剂,与前体化合物的重链中4个氨基酸的差异,依库珠单抗(ECUL)。
■首先,利用质谱(MS)来表征RAVUL并验证与其前身的差异,第二,验证并实施RAVUL的实验室开发测试(LDT),以进行定量治疗监测。
■使用飞行时间质谱仪(TOF-MS)通过消化和还原实验来表征和区分RAVUL和ECUL之间的分子量差异。并行,在高通量OrbitrapMS平台上,利用具有轻链检测和定量的IgG4富集对RAVUL的LDT进行了验证和实施.
■TOF-MS平台允许验证RAVUL和ECUL之间的质量差异,并为新的完整蛋白质定量软件提供概念证明。使用完整的轻链定量对轨道阱MS上的LDT进行了验证和实施,具有不能区分ECUL和RAVUL的限制。LDT的分析测量范围从5到600mcg/mL,测定间的不精确性≤13%CV(n=13)和准确度与预期值误差<4%(n=20)。
■TOF-MS是一种多功能的开发平台,可用于表征和验证ECUL和RAVUL重链之间的分子量差异。使用orbitrap-MS进行RAVUL的常规实验室测试是可行的,以使用完整轻链的质量进行定量。这两个平台,合并,为临床实验室LDTs的开发提供了无与伦比的价值。
UNASSIGNED: Ravulizumab (RAVUL) is a new complement inhibitor, with a difference of 4 amino acids in the heavy chain from a predecessor compound, eculizumab (ECUL).
UNASSIGNED: First, to utilize mass spectrometry (MS) to characterize RAVUL and verify differences from its predecessor and, second, to validate and implement a lab developed test (LDT) for RAVUL that will allow for quantitative therapeutic monitoring.
UNASSIGNED: A time-of-flight mass spectrometer (TOF-MS) was used to characterize and differentiate the molecular weight differences between RAVUL and ECUL by both digest and reduction experiments. In parallel, an LDT for RAVUL was validated and implemented utilizing IgG4 enrichment with light chain detection and quantitation on a high throughput orbitrap MS platform.
UNASSIGNED: The TOF-MS platform allowed for the mass difference between RAVUL and ECUL to be verified along with providing a proof of concept for a new intact protein quantitation software. An LDT on an orbitrap MS was validated and implemented using intact light chain quantitation, with the limitation that it cannot differentiate between ECUL and RAVUL. The LDT has an analytical measuring range from 5 to 600 mcg/mL, inter-assay imprecision of ≤13% CV (n = 13) and accuracy with <4% error from expected values (n = 20).
UNASSIGNED: The TOF-MS is a versatile development platform that can be used to characterize and verify the molecular weight differences between the ECUL and RAVUL heavy chains. Routine laboratory testing for RAVUL was viable using an orbitrap-MS to quantitate using the mass of the intact light chain. These two platforms, combined, provide incomparable value in development of LDTs for the clinical laboratory.