Ondigo Bartholomew N, Park Gregory S, Ayieko Cyrus, Nyangahu Donald D, Wasswa Ronald, John Chandy C
Department of Biochemistry and Molecular Biology, Egerton University, Nakuru, Kenya.
Centre for Global Health Research, Kenya Medical Research Institute, Kisumu, Kenya.
PeerJ. 2019 Jan 3;7:e6120. doi: 10.7717/peerj.6120. eCollection 2019.
New reagents have emerged allowing researchers to assess a growing number of vaccine-associated immune parameters. Multiplex immunoassay(s) are emerging as efficient high-throughput assays in malaria serology. Currently, commercial vendors market several bead reagents for cytometric bead assays (CBA) but relative performances are not well published. We have compared two types of bead-based multiplex assays to measure relative antibody levels to malarial antigens.
Assays for the measurement of antibodies to five vaccine candidates using non-magnetic and magnetic fluorescent microspheres were compared for their performances with a Bio-Plex instrument. Mean fluorescence intensity (MFI) was determined from individuals from western Kenya and compared to known positive and negative control plasma samples.
recombinant antigens were successfully coupled to both non-magnetic and magnetic beads in multiplex assays. MFIs between the two bead types were comparable for all antigens tested. Bead recovery was superior with magnetic beads for all antigens. MFI values of stored non-magnetic coupled beads did not differ from freshly coupled beads, though they showed higher levels of bead aggregation.
Magnetic and non-magnetic beads performed similarly in antibody assays. Magnetic beads were more expensive, but had higher bead recovery, were more convenient to use, and provided rapid and easy protocol manipulation. Magnetic beads are a suitable alternative to non-magnetic beads in malarial antibody serology.
新试剂不断涌现，使研究人员能够评估越来越多与疫苗相关的免疫参数。多重免疫测定正在成为疟疾血清学中高效的高通量检测方法。目前，商业供应商销售几种用于细胞计数珠分析（CBA）的微珠试剂，但相对性能尚未得到充分报道。我们比较了两种基于微珠的多重分析方法，以测量针对疟疾抗原的相对抗体水平。
使用非磁性和磁性荧光微球对五种疫苗候选物的抗体测量分析方法，通过Bio-Plex仪器比较其性能。从肯尼亚西部的个体中测定平均荧光强度（MFI），并与已知的阳性和阴性对照血浆样本进行比较。
在多重分析中，重组抗原成功地与非磁性和磁性微球偶联。对于所有测试抗原，两种微珠类型之间的MFI相当。对于所有抗原，磁性微球的微珠回收率更高。储存的非磁性偶联微球的MFI值与新鲜偶联的微球没有差异，尽管它们显示出更高水平的微球聚集。
磁性和非磁性微球在抗体分析中的表现相似。磁性微球更昂贵，但微珠回收率更高，使用更方便，并且提供快速简便的方案操作。在疟疾抗体血清学中，磁性微球是替代非磁性微球的合适选择。
