Institute for Reference Materials and Measurements (IRMM), Joint Research Centre, European Commission, Geel, Belgium.
Clin Chem Lab Med. 2010 Jun;48(6):805-13. doi: 10.1515/CCLM.2010.166.
The availability of matrix reference materials is essential for the standardisation of (immuno)assays used to measure proteins. The reference material ERM-DA470 (previously called CRM470) certified in 1993 has led to a large degree of harmonisation of these assays. A new serum protein reference material has now been produced (ERM-DA470k). It is intended to replace ERM-DA470, and will additionally be certified for beta(2)-microglobulin (B2M).
Serum from 390 healthy donors was pooled and processed so as to stabilise, delipidate and 'maturate' it. Purified C-reactive protein (CRP) and recombinant B2M were added. Pilot batches were produced to study the stability, homogeneity, and commutability of the material. On the basis of the results with the trial batches it was decided to proceed with the processing of the main batch of a candidate reference material.
Two pilot batches were produced and the processed and spiked serum lyophilised after filling (1 mL). The B2M in the material was shown to be stable and commutable. For CRP, it was discovered that freeze-drying led to a decrease in measurable protein. The main batch of candidate reference material was produced and fulfilled the required criteria in terms of optical transparency, homogeneity and stability.
A new serum protein reference material has been produced with the properties required for a serum protein reference material for 14 proteins. An apparent loss of CRP of approximately 20% was observed upon freeze-drying of the material.
基质参考物质的可用性对于标准化用于测量蛋白质的（免疫）测定法至关重要。1993 年认证的参考物质 ERM-DA470（以前称为 CRM470）在很大程度上促成了这些测定法的协调一致。现在已经生产出一种新的血清蛋白参考物质（ERM-DA470k）。它旨在替代 ERM-DA470，并将另外针对β(2)-微球蛋白（B2M）进行认证。
从 390 名健康供体中采集血清，然后进行混合处理以使其稳定、去脂和“成熟”。添加了纯化的 C 反应蛋白（CRP）和重组 B2M。生产了试验批来研究材料的稳定性、均一性和可互换性。根据试验批的结果，决定继续处理候选参考物质的主要批次。
生产了两个试验批，并在填充后（1 毫升）冻干处理和添加了 B2M 的血清。该材料中的 B2M 表现出稳定性和可互换性。对于 CRP，发现冻干会导致可测量蛋白的减少。生产了候选参考物质的主要批次，并在光学透明度、均一性和稳定性方面符合要求。
已经生产出一种具有 14 种血清蛋白参考物质所需特性的新的血清蛋白参考物质。在冻干该材料时，观察到 CRP 明显损失约 20%。
